Biochemical reaction methods and reagents comprising intrinsically disordered regions

Abstract

20 26 23 41 01 18 S ep 2 02 6 B I O C H E M I C A L R E A C T I O N M E T H O D S A N D R E A G E N T S C O M P R I S I N G 2 0 2 6 2 3 4 1 0 1 1 8 2 0 2 6 I N T R I N S I C A L L Y D I S O R D E R E D R E G I O N S S e p A B S T R A C T

Description

This is a divisional of Australian Patent Application No. 2020382121, the entire contents of which are incorporated herein by reference. 5 FIELD OF THE INVENTION The invention relates to processes for performing biochemical reactions, such as in an aqueous in vitro reaction system. The processes involve macromolecules, 10 particularly polypeptides, comprising one or more functional intrinsically disordered regions (IDRs). The invention also relates to IDR-macromolecules, including IDR-polypeptides, including macromolecules or polypeptides comprising a tagged amino acid sequence which comprises or consists of one or more functional IDRs. Such functional IDRs are capable of increasing the efficiency of the biochemical reaction. 15 The invention relates to kits comprising any such macromolecules and polypeptides. The invention further relates to processes for stimulating or enhancing liquid-liquid demixing in a solution using any such macromolecules and polypeptides, including in combination with multivalent metal ions, thereby providing reagents capable of increasing the efficiency of a biochemical reaction. 20 BACKGROUND TO THE INVENTION The performance of biochemical reactions, and in particular in vitro biochemical reactions, is of fundamental importance in the biological sciences. Many biochemical 25 reactions may need to be performed outside of the laboratory, such as at the point of care or in the field. In these settings it may not be possible to control biochemical reactions in the precise manner afforded by the laboratory environment. Improving the efficiency of biochemical reactions performed in these settings would be of value. Indeed, it may be desirable to increase the efficiency of biochemical reactions, 30 regardless of the exact setting, including in vitro and in vivo biochemical reactions. The present invention addresses these issues. Many biochemical reactions require the use of co-factors to aid in driving performance efficiency. One particular example of such a co-factor is a 2026234101   18 Sep 2026 macromolecular crowding agent. Crowding agents are essential for the performance of many biochemical reactions. A notable example is the recombinase polymerase amplification (RPA) system for the amplification of nucleic acids. The use of a crowding agent has been considered essential in driving RPA performance efficiency. 5 However, crowding agents may have drawbacks. Accordingly, alternative means for driving performance efficiency of biochemical reactions, including RPA, and that obviate the need for added / exogenous crowding agents would be of use. In addition, reagents that add to or synergise with the functional effects of crowding agents in increasing the performance efficiency of biochemical reactions would be of use. The 10 present invention also addresses these issues. SUMMARY OF THE INVENTION The present invention provides a process of performing a biochemical reaction 15 in an aqueous in vitro reaction system, wherein the biochemical reaction is dependent on the function of at least one reaction macromolecule, optionally at least one reaction polypeptide, the process comprising: introducing at least one IDR-macromolecule into the in vitro reaction system under conditions suitable for performing the reaction, wherein the at least one IDR-macromolecule comprises one or more functional 20 intrinsically disordered regions (IDRs), wherein upon introduction of the at least one IDR-macromolecule into the in vitro reaction system the efficiency of the biochemical reaction is increased by the at least one IDR-macromolecule; preferably wherein the at least one IDR-macromolecule is at least one IDR-polypeptide. In the above-described process, the biochemical reaction may be dependent on 25 the function of the at least one IDR-macromolecule, optionally the at least one IDR-polypeptide, wherein upon its introduction into the in vitro reaction system the at least one IDR-macromolecule or the at least one IDR-polypeptide performs its reaction function in the biochemical reaction and increases the efficiency of the reaction. Any of the herein-described processes may further comprise maintaining the 30 IDR-macromolecule or the IDR-polypeptide in the system to cause liquid-liquid demixing and the formation of a plurality of phase-separated aqueous compartments 2026234101   18 Sep 2026 within the system by the IDR-macromolecule or the IDR-polypeptide, thereby increasing the efficiency of the biochemical reaction in the system. Any of the herein-described processes may further comprise maintaining the IDR-macromolecule or the IDR-polypeptide in the system to cause molecules necessary 5 for the performance of the reaction to co-localise with the IDR-macromolecule or the IDR-polypeptide within the plurality of phase-separated aqueous compartments, or to further stimulate or enhance co-localisation of molecules necessary for the performance of the reaction with the IDR-macromolecule or the IDR-polypeptide within the plurality of phase-separated aqueous compartments, thereby increasing the efficiency of the 10 biochemical reaction in the system. In any of the herein -described processes the plurality of phase-separated aqueous compartments may be a plurality of detectable phase-separated aqueous particles. In an additional aspect the invention provides a process of performing a 15   biochemical reaction in an aqueous in vitro reaction system, wherein the biochemical reaction is dependent on the function of at least one reaction macromolecule, optionally at least one reaction polypeptide, the process comprising: introducing at least one polypeptide tagged with an amino acid sequence comprising or consisting of one or more functional intrinsically disordered regions (IDRs) (IDR-polypeptide) into the in 20 vitro reaction system under conditions suitable for performing the reaction, and maintaining the IDR-polypeptide in the system to cause liquid-liquid demixing and the formation of a plurality of phase-separated aqueous compartments, preferably detectible phase-separated aqueous particles, within the system by the IDR-polypeptide and to cause molecules necessary for the performance of the reaction to co-localise with the 25 IDR-polypeptide within the compartments thereby increasing the efficiency of the biochemical reaction in the system. Optionally, in a process according to this additional aspect, the biochemical reaction is dependent on the function of at least one reaction polypeptide, wherein the reaction polypeptide is the at least one IDR-polypeptide, wherein upon introduction into 30 the system the at least one IDR-polypeptide performs its reaction function in the biochemical reaction and increases the efficiency of the reaction in the system. 2026234101   18 Sep 2026 In any of the processes according to this additional aspect, the conditions suitable for performing the reaction may further comprise providing multivalent metal ions to the IDR-polypeptide thereby further stimulating or enhancing the liquid-liquid demixing and the formation of the plurality of phase-separated aqueous compartments 5 caused by the IDR-polypeptide and thereby further increasing the efficiency of the biochemical reaction in the system; optionally wherein the multivalent metal ions are provided at a concentration of about 22 mM or more, preferably wherein the multivalent metal ions are provided at a concentration of between about 22 mM to 50 mM. The multivalent metal ions may be divalent metal ions, optionally Mg2+, Mn2+, Ca2+, Co2+ or 10   Ni2+, preferably Mg2+, Mn2+ or Ca2+, more preferably Mg2+. In any of the processes according to this additional aspect, the conditions suitable for performing the reaction may further comprise providing ATP to the IDR-polypeptide in the in vitro reaction system, thereby further simulating or enhancing the liquid-liquid demixing and the formation of the plurality of phase-separated aqueous 15 compartments caused by the IDR-polypeptide and thereby further increasing the efficiency of the biochemical reaction in the system, wherein ATP is provided in the system at a concentration of ImM to 3.5mM, optionally ImM to 2mM, preferably ImM. In any of the processes according to this additional aspect, the conditions 20 suitable for performing the reaction may further comprise providing multivalent metal ions to the IDR-polypeptide thereby further stimulating or enhancing molecules necessary for the performance of the reaction to co-localise with the IDR-polypeptide within the plurality of phase-separated aqueous compartments thereby further increasing the efficiency of the biochemical reaction in the system; optionally wherein the 25 multivalent metal ions are provided at a concentration of about 22 mM or more, preferably wherein the multivalent metal ions are provided at a concentration of between about 22 mM to 50 mM. The multivalent metal ions may be divalent metal ions, optionally Mg2+, Mn2+, Ca2+, Co2+ or Ni2+, preferably Mg2+, Mn2+ or Ca2+, more preferably Mg2+. 30           In any of the processes according to this additional aspect, the conditions suitable for performing the reaction may further comprise providing ATP to the IDR- 2026234101   18 Sep 2026 polypeptide in the in vitro reaction system, thereby further stimulating or enhancing molecules necessary for the performance of the reaction to co-localise with the IDR-polypeptide within the plurality of phase-separated aqueous compartments and thereby further increasing the efficiency of the biochemical reaction in the system, wherein ATP 5 is provided in the system at a concentration of ImM to 3.5mM, optionally ImM to 2mM, preferably ImM. In any of the processes according to this additional aspect, the efficiency of the reaction in the system may be increased by the IDR-polypeptide compared to the efficiency of the reaction in the system following introduction of the at least one 10   polypeptide under the same reaction conditions except that the at least one polypeptide has not been tagged with the amino acid sequence comprising or consisting of the one or more functional IDRs. The invention also provides a process of performing a biochemical reaction in an aqueous in vitro reaction system, wherein the biochemical reaction is dependent on 15   the function of at least one reaction macromolecule, optionally at least one reaction polypeptide, the process comprising: i. introducing molecules comprising at least one IDR-macromolecule into the system under conditions suitable for performing the reaction, wherein the at 20          least one IDR-macromolecule comprises one or more functional intrinsically disordered regions (IDRs), preferably wherein the at least one IDR-macromolecule is at least one IDR-polypeptide; ii. maintaining the IDR-macromolecule or the IDR-polypeptide in the system to 25          cause liquid-liquid demixing in the system, wherein the liquid-liquid demixing is caused by the IDR-macromolecule or the IDR-polypeptide and forms a plurality of phase-separated aqueous compartments within the system; iii. maintaining the IDR-macromolecule or the IDR-polypeptide in the system to 30          cause molecules necessary for the performance of the reaction to co-localise 2026234101   18 Sep 2026 with the IDR-macromolecule or the IDR-polypeptide within the compartments; and iv. allowing the biochemical reaction to proceed within the compartments; wherein 5          the efficiency of the biochemical reaction in the system is increased by the presence of the at least one IDR-macromolecule. In the above-described process, the biochemical reaction may be dependent on the function of the at least one IDR-macromolecule, optionally the at least one IDR-polypeptide, wherein upon its introduction into the in vitro reaction system the at least 10 one IDR-macromolecule or the at least one IDR-polypeptide performs its reaction function in the biochemical reaction and increases the efficiency of the reaction. The plurality of phase-separated aqueous compartments may be a plurality of detectable phase-separated aqueous particles. In a further aspect the invention provides a process of performing a biochemical 15   reaction in an aqueous in vitro reaction system, wherein the biochemical reaction is dependent on the function of at least one reaction macromolecule, optionally at least one reaction polypeptide, the process comprising: i. introducing molecules comprising at least one polypeptide tagged with an amino 20          acid sequence comprising or consisting of one or more functional intrinsically disordered regions (IDRs) (IDR-polypeptide) into the system under conditions suitable for performing the reaction; ii. maintaining the IDR-polypeptide in the system to cause liquid-liquid demixing 25          and the formation of a plurality of phase-separated aqueous compartments, preferably detectible phase-separated aqueous particles, within the system, wherein the liquid-liquid demixing is caused by the IDR-polypeptide; iii. maintaining the IDR-polypeptide in the system to cause molecules necessary for 30          the performance of the reaction to co-localise with the IDR-polypeptide within the compartments; and 2026234101   18 Sep 2026 iv. allowing the biochemical reaction to proceed within the compartments; wherein the efficiency of the biochemical reaction in the system is increased by the presence of the at least one IDR- polypeptide. Optionally, in a process according to this further aspect, the biochemical 5 reaction is dependent on the function of at least one reaction polypeptide, wherein the reaction polypeptide is the at least one IDR-polypeptide, wherein upon introduction into the system the at least one IDR-polypeptide performs its reaction function in the biochemical reaction and increases the efficiency of the reaction in the system. In any of the processes according to this further aspect, the conditions suitable 10   for performing the reaction may further comprise providing multivalent metal ions to the IDR-polypeptide thereby further stimulating or enhancing the liquid-liquid demixing and the formation of the plurality of phase-separated aqueous compartments caused by the IDR-polypeptide and thereby further increasing the efficiency of the biochemical reaction in the system; optionally wherein the multivalent metal ions are provided at a 15   concentration of about 22 mM or more, preferably wherein the multivalent metal ions are provided at a concentration of between about 22 mM to 50 mM. The multivalent metal ions may be divalent metal ions, optionally Mg2+, Mn2+, Ca2+, Co2+ or Ni2+, preferably Mg2+, Mn2+ or Ca2+, more preferably Mg2+. In any of the processes according to this further aspect, the conditions suitable 20 for performing the reaction may further comprise providing ATP to the IDR-polypeptide in the in vitro reaction system, thereby further simulating or enhancing the liquid-liquid demixing and the formation of the plurality of phase-separated aqueous compartments caused by the IDR-polypeptide and thereby further increasing the efficiency of the biochemical reaction in the system, wherein ATP is provided in the 25 system at a concentration of ImM to 3.5mM, optionally ImM to 2mM, preferably ImM. In any of the processes according to this further aspect, the conditions suitable for performing the reaction may further comprise providing multivalent metal ions to the IDR-polypeptide thereby further stimulating or enhancing molecules necessary for 30 the performance of the reaction to co-localise with the IDR-polypeptide within the plurality of phase-separated aqueous compartments thereby further increasing the 2026234101   18 Sep 2026 efficiency of the biochemical reaction in the system; optionally wherein the multivalent metal ions are provided at a concentration of about 22 mM or more, preferably wherein the multivalent metal ions are provided at a concentration of between about 22 mM to 50 mM. The multivalent metal ions maybe divalent metal ions, optionally Mg2+, Mn2+, 5   Ca2+, Co2+ or Ni2+, preferably Mg2+, Mn2+ or Ca2+, more preferably Mg2+. In any of the processes according to this further aspect, the conditions suitable for performing the reaction may further comprise providing ATP to the IDR-macromolecule or IDR-polypeptide in the in vitro reaction system, thereby further stimulating or enhancing molecules necessary for the performance of the reaction to co-10   localise with the IDR-polypeptide within the plurality of phase-separated aqueous compartments and thereby further increasing the efficiency of the biochemical reaction in the system, wherein ATP is provided in the system at a concentration of ImM to 3.5mM, optionally ImM to 2mM, preferably ImM. In any of the processes according to this further aspect, the conditions suitable 15   for performing the reaction may further comprise providing multivalent metal ions to the IDR-polypeptide thereby further stimulating or enhancing molecules necessary for the performance of the reaction to co-localise with the IDR-macromolecule or the IDR-polypeptide within the plurality of phase-separated aqueous compartments thereby further increasing the efficiency of the biochemical reaction in the system; optionally 20 wherein the multivalent metal ions are provided at a concentration of about 22 mM or more, preferably wherein the multivalent metal ions are provided at a concentration of between about 22 mM to 50 mM. The multivalent metal ions may be divalent metal ions, optionally Mg2+, Mn2+, Ca2+, Co2+ or Ni2+, preferably Mg2+, Mn2+ or Ca2+, more preferably Mg2+. 25           In any of the processes according to this further aspect, the efficiency of the reaction in the system may be increased by the IDR-polypeptide compared to the efficiency of the reaction in the system following introduction of the at least one polypeptide under the same reaction conditions except that the at least one polypeptide has not been tagged with the amino acid sequence comprising or consisting of the one 30 or more functional IDRs. 2026234101   18 Sep 2026 In any of the above-described processes, the process may be a biochemical reaction for synthesizing a nucleic acid molecule in an in vitro reaction system comprising: (a) providing at least one nucleic acid primer; 5           (b) providing a target nucleic acid molecule comprising at least one target strand, and contacting the at least one nucleic acid primer with the target strand, thereby forming a double stranded structure; (c) providing the IDR-macromolecule as an IDR-polypeptide, wherein the IDR-polypeptide is polymerase or one or more polypeptide co-factors; and 10           (d) allowing the reaction to proceed, thereby extending the 3 ’ end of the at least one nucleic acid primer with polymerase and dNTPs, optionally in the presence of one or more polypeptide co-factors, to generate a double stranded nucleic acid, wherein the first strand comprises a sequence of the target strand and the second strand comprises a sequence which is 15             complementary thereto. Alternatively, in any of the above-described processes, the process may be a biochemical for amplifying a single stranded target nucleic acid molecule or a double stranded target nucleic acid molecule in an in vitro reaction system, preferably wherein the target nucleic acid molecule is a DNA molecule. 20          The process may be a biochemical reaction for amplifying a double stranded target nucleic acid molecule in an in vitro reaction system comprising: (a) providing first and second nucleic acid primers; (b) providing a double stranded target nucleic acid molecule comprising a first strand and a second strand, and contacting the first and second nucleic acid 25              primers with the target nucleic acid molecule thereby forming a first double stranded structure with the first strand and a second double stranded structure with the second strand; (c) providing the IDR-macromolecule as an IDR-polypeptide, wherein the IDR-polypeptide is polymerase or one or more protein co-factors; 30           (d) allowing the reaction to proceed, thereby extending the 3’ ends of the first and second nucleic acid primers with polymerase and dNTPs, optionally in 2026234101   18 Sep 2026 the presence of one or more protein co-factors, to generate first and second double stranded nucleic acids; and (e) repeating steps (b) to (d) until a desired degree of amplification is reached. In the above-described process for amplifying a double stranded target nucleic 5 acid molecule in an in vitro reaction system, the process may be a recombinase polymerase amplification (RPA) process of amplifying the double stranded target nucleic acid molecule in the in vitro reaction system comprising: (a) providing a recombinase agent, optionally a recombinase loading protein, a single strand stabilizing agent, polymerase, first and a second nucleic acid 10              primers, double stranded target nucleic acid comprising a first strand and a second strand, and optionally an exonuclease such as Exonuclease III; (b) contacting the recombinase agent with the first and second nucleic acid primers and optionally with the recombinase loading protein to form first and second nucleoprotein primers which comprise a single stranded region at 15               their 3’ends; (c) contacting the first and second nucleoprotein primers with the target nucleic acid molecule thereby forming a first double stranded structure with the first strand and a second double stranded structure with the second strand; (d) allowing the reaction to proceed, thereby extending the 3 ’ end of the first 20             and second nucleoprotein primers with polymerase and dNTPs to generate first and second double stranded nucleic acids and first and a second displaced nucleic acid strands, wherein the single strand stabilizing agent stabilizes the first and second displaced strands; and (e) continuing the reaction by repeating steps (b) to (d) until a desired degree of 25               amplification is reached; wherein the recombinase agent, and / or the recombinase loading protein, and / or the single strand stabilizing agent, and / or the polymerase is provided as the IDR-polypeptide. In the above-described RPA process of amplifying a double stranded target 30 nucleic acid molecule in the in vitro reaction system, the recombinase agent may be selected from the group consisting of UvsX, T4 UvsX, T6 UvsX, RBI8 UvsX, E.coli 2026234101   18 Sep 2026 phage wV7 UvsX, Shigella phage CB8 UvsX, Shigella phage Shfl2 UvsX, E.coli phage ARI UvsX, phage vB_EcoM_G4507 UvsX, Shigella phage SHFML-11 UvsX, Escherichia phage vB_EcoM_DalCa UvsX, E. coli RecA, E. coli RadA, E. coli RadB, E. coli Rad 51 or any functional analog, homolog or derivative thereof, and any 5 combination thereof, preferably wherein the recombinase agent is UvsX, more preferably Escherichia phage vBEcoMDalCa UvsX. In any one of the above-described RPA processes of amplifying a double stranded target nucleic acid molecule in the in vitro reaction system, the process may include a recombinase loading protein and wherein the recombinase loading protein is 10 selected from the group consisting of UvsY, E. coli RecO, E. coli RecR or any functional analog, homolog or derivative thereof, and any combination thereof, preferably wherein the recombinase loading protein is UvsY, more preferably Escherichia phage STO UvsY. In any one of the above-described RPA processes of amplifying a double 15 stranded target nucleic acid molecule in the in vitro reaction system, the polymerase may be a eukaryotic polymerase selected from the group consisting of pol-a, pol-P, pol-6, pol-c or any functional analog, homolog or derivative thereof, and any combination thereof. The polymerase may be a prokaryotic polymerase selected from the group consisting of Bacillus stearothermophilus polymerase I large fragment, Bacillus subtilis 20 Pol I large fragment (Bsu polymerase), Listeria monocytogenes DNA polymerase I, S. aureus DNA polymerase I (Sau polymerase), E. coli DNA polymerase I Klenow fragment, E. coli DNA polymerase I, E. coli DNA polymerase II, E. coli DNA polymerase III, E. coli DNA polymerase IV, E. coli DNA polymerase V, or any functional analog, homolog or derivative thereof, and any combination thereof, 25 preferably wherein the polymerase is S. aureus DNA polymerase I (Sau polymerase) or Bacillus subtilis Pol I large fragment (Bsu polymerase). The polymerase may be a bacteriophage polymerase selected from the group consisting of bacteriophage T4 gp43 DNA polymerase, T7 DNA polymerase and Phi-29 DNA polymerase, or any functional analog, homolog or derivative thereof, and any combination thereof. 30          In any one of the above-described RPA processes of amplifying a double stranded target nucleic acid molecule in the in vitro reaction system, the single strand 2026234101   18 Sep 2026 stabilizing agent may be selected from the group consisting of Gp32, E. coli SSB protein, phage T4 Gp32 protein, phage Rb69 Gp32, phage vB_EcoM_NBGl Gp32, or any functional analog, homolog or derivative thereof, and any combination thereof, preferably the single strand stabilizing agent is Gp32 or phage vB_EcoM_NBGl Gp32. 5          In any one of the above-described RPA processes of amplifying a double stranded target nucleic acid molecule in the in vitro reaction system, only the recombinase agent may be provided as the IDR-polypeptide, or only the recombinase loading protein may be provided as the IDR-polypeptide, or only the single strand stabilizing agent may be provided as the IDR-polypeptide, or only the polymerase may 10 be provided as the IDR-polypeptide, or only the exonuclease may be provided as the IDR-polypeptide. In any one of the above-described RPA processes of amplifying a double stranded target nucleic acid molecule in the in vitro reaction system, the one or more functional IDRs of the IDR-polypeptide may be tagged to the IDR-polypeptide as an 15   amino acid sequence comprising or consisting of the one or more IDRs such that the IDR-polypeptide is a genetically engineered fusion protein, wherein the one or more functional IDRs are located at the C-terminus of the IDR-polypeptide, at the N-terminus of the IDR-polypeptide, or at both the C-terminus of the IDR-polypeptide and the N-terminus of the IDR-polypeptide, or at any amino acid position along the length of the 20 polypeptide. In any one of the above-described processes, the one or more functional IDRs of the IDR-macromolecule or the IDR-polypeptide may be characterised as a sequence of amino acids which scores greater than 0.5 when analysed by the algorithm MetaDisorder. 25          In any one of the above-described processes, the one or more functional IDRs of the IDR-macromolecule or the IDR-polypeptide may comprise or consist of an amino acid sequence which includes one or more repeats of the tri-peptide sequence RGG. In any such process, the one or more functional IDRs of the IDR-macromolecule or the IDR-polypeptide may comprise or consist of an amino acid sequence which further 30 includes one or more repeats of the di-peptide sequence FG. In any such process, the one or more functional IDRs of the IDR-macromolecule or the IDR-polypeptide may 2026234101   18 Sep 2026 comprise or consist of an amino acid sequence which further includes at least one aromatic amino acid residue consisting of tyrosine or phenylalanine. In any one of the above-described processes, the one or more functional IDRs of the IDR-macromolecule or the IDR-polypeptide may comprise or consist of the amino 5 acid sequence of i. (YNPQGGYQQ)n(SEQ ID NO: 19), wherein n is a positive integer between 1 and 10, optionally wherein n = 1, 2, or 3; or ii. (YSPTSPS)n (SEQ ID NO: 124), wherein n is a positive integer between 1 and 10, optionally wherein n = 1, 2, or 3; or 10 iii. (FSPTSPT)n(SEQ ID NO: 125), wherein n is a positive integer between 1 and 10, optionally wherein n = 1, 2, or 3; or iv. (YSPTSP-A / N / G)n (SEQ ID NO: 126), wherein n is a positive integer between 1 and 10, optionally wherein n = 1, 2, or 3; or v. (YSPGSPA)n (SEQ ID NO: 127), wherein n is a positive integer between 1 and 15            10, optionally wherein n = 1, 2, or 3. In any one of the above-described processes, the one or more functional IDRs of the IDR-macromolecule or the IDR-polypeptide may comprise or consist of an amino acid sequence which is glutamine rich, optionally wherein the amino acid sequence comprises at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 20 consecutive glutamine residues. In any such process, the one or more functional IDRs of the IDR-macromolecule or the IDR-polypeptide may comprise or consist of an amino acid sequence comprising one or more repeats of the tri-peptide sequence QQQ. In any such process, the one or more functional IDRs of the IDR-macromolecule or the IDR-polypeptide comprises or consists of the amino acid sequence of (QQQPQY)n (SEQ ID 25 NO: 128), wherein n is a positive integer between 1 and 10, optionally wherein n = 1,2, or 3. In any one of the above-described processes, the one or more functional IDRs of the IDR-macromolecule or the IDR-polypeptide may comprise or consist of a sequence of at least 5 consecutive amino acids of SEQ ID NO: 1. 2026234101   18 Sep 2026 In any one of the above-described processes, the one or more functional IDRs of the IDR-macromolecule or the IDR-polypeptide may comprise or consist of an amino acid sequence of at least 5 consecutive amino acids of SEQ ID NO:9. In any one of the above-described processes, the one or more functional IDRs of 5 the IDR-macromolecule or the IDR-polypeptide may comprise an amino acid sequence containing one or more aromatic tyrosine residues and one or more phenylalanine residues which can engage in aromatic cation-pi interactions with multivalent metal ions, preferably divalent metal ions. In any one of the above-described processes, the one or more functional IDRs of 10   the IDR-macromolecule or the IDR-polypeptide may comprise an amino acid sequence containing one or more arginine residues which can engage in guanidine-metal interactions with multivalent metal ions, preferably divalent metal ions. In any one of the above-described processes, the IDR-macromolecule or the IDR-polypeptide may comprise or consist of the macromolecule or polypeptide tagged 15   with an amino acid sequence which comprises or consists of the amino acid sequence of any one of SEQ ID NOs 1 to 43, or which comprises or consists of a functional variant amino acid sequence of SEQ ID NOs 1 to 43, e.g. which has 80% or more identity to any one of SEQ ID NOs 1 to 43. In any one of the above-described RPA processes of amplifying a double 20 stranded target nucleic acid molecule in the in vitro reaction system, the IDR-polypeptide may be a single strand stabilizing agent which is Gp32 and which has the amino acid sequence of any one of SEQ ID NOs 65 to 88, or wherein the IDR-polypeptide is a functional variant thereof, e.g. an IDR-polypeptide having an amino acid sequence which has 80% or more identity to any one of SEQ ID NOs 65 to 88. 25          In any one of the above-described RPA processes of amplifying a double stranded target nucleic acid molecule in the in vitro reaction system, the IDR-polypeptide may be a recombinase agent which is UvsX and which has the amino acid sequence of any one of SEQ ID NOs 44 to 59, or wherein the IDR-polypeptide is a functional variant thereof, e.g. an IDR-polypeptide having an amino acid sequence 30 which has 80% or more identity to any one of SEQ ID NOs 44 to 59. 2026234101   18 Sep 2026 In any one of the above-described RPA processes of amplifying a double stranded target nucleic acid molecule in the in vitro reaction system, the IDR-polypeptide may be a recombinase loading protein which is UvsY and which has the amino acid sequence of any one of SEQ ID NOs 60 to 64, or wherein the IDR- 5 polypeptide is a functional variant thereof, e.g. an IDR-polypeptide having an amino acid sequence which has 80% or more identity to any one of SEQ ID NOs 60 to 64. In any one of the above-described processes, the process may further comprise providing multivalent metal ions to the IDR-macromolecule or IDR-polypeptide in the in vitro reaction system, thereby further simulating or enhancing the liquid-liquid 10   demixing in the in vitro reaction system, thereby further increasing the efficiency of the biochemical reaction in the system,wherein the multivalent metal ions further stimulate or enhance the formation of the plurality of phase-separated aqueous compartments within the system thereby farther increasing the efficiency of the biochemical reaction in the system, preferably wherein the multivalent metal ions further stimulate or 15 enhance the formation of a plurality of detectable phase-separated aqueous particles; optionally wherein the multivalent metal ions are provided at a concentration of; optionally wherein the multivalent metal ions are provided at a concentration of about 22 mM or more, preferably wherein the multivalent metal ions are provided at a concentration of between about 22 mM to 50 mM. In any such process, the multivalent 20 metal ions may be divalent metal ions, optionally Mg2+, Mn2+, Ca2+, Co2+ or Ni2+, preferably Mg2+, Mn2+ or Ca2+, more preferably Mg2+. In any one of the above-described processes, the conditions suitable for performing the reaction may further comprise providing ATP to the IDR-macromolecule or the IDR-polypeptide in the in vitro reaction system, thereby further 25 simulating or enhancing the liquid-liquid demixing and the formation of the plurality of phase-separated aqueous compartments caused by the IDR-macromolecule or the IDR-polypeptide and thereby further increasing the efficiency of the biochemical reaction in the system, wherein ATP is provided in the system at a concentration of ImM to 3.5mM, optionally ImM to 2mM, preferably ImM. 30          In any one of the above-described processes, the conditions suitable for performing the reaction may further comprise providing multivalent metal ions to the 2026234101   18 Sep 2026 IDR-polypeptide thereby further stimulating or enhancing molecules necessary for the performance of the reaction to co-localise with the IDR-macromolecule or the IDR-polypeptide within the plurality of phase-separated aqueous compartments thereby further increasing the efficiency of the biochemical reaction in the system; optionally 5 wherein the multivalent metal ions are provided at a concentration of about 22 mM or more, preferably wherein the multivalent metal ions are provided at a concentration of between about 22 mM to 50 mM. The multivalent metal ions may be divalent metal ions, optionally Mg2+, Mn2+, Ca2+, Co2+ or Ni2+, preferably Mg2+, Mn2+ or Ca2+, more preferably Mg2+. 10          In any one of the above-described processes, the conditions suitable for performing the reaction may further comprise providing ATP to the IDR-macromolecule or IDR-polypeptide in the in vitro reaction system, thereby further stimulating or enhancing molecules necessary for the performance of the reaction to colocalise with the IDR-polypeptide within the plurality of phase-separated aqueous 15 compartments and thereby further increasing the efficiency of the biochemical reaction in the system, wherein ATP is provided in the system at a concentration of ImM to 3.5mM, optionally ImM to 2mM, preferably ImM. In any one of the above-described processes, the biochemical reaction may be performed in a solid phase reaction system which comprises a surface. In any such 20 process the biochemical reaction may be a process of amplifying a single stranded target nucleic acid molecule or a double stranded target nucleic acid molecule in the in vitro reaction system as described above, wherein the at least one nucleic acid primer and / or the IDR-macromolecule and / or the one or more polypeptide co-factors are attached to the surface. 25          In any one of the above-described RPA processes of amplifying a double stranded target nucleic acid molecule in the in vitro reaction system, the reaction may be performed in a solid phase reaction system which comprises a surface, and wherein the recombinase agent and / or the recombinase loading protein and / or the single strand stabilizing agent and / or the polymerase and / or the exonuclease and / or the first nucleic 30 acid primer and / or the second nucleic acid primer is attached to the surface, preferably wherein: (i) the first nucleic acid primer or the second nucleic acid primer is attached to 2026234101   18 Sep 2026 the surface; or (ii) both the first and second nucleic acid primers are attached to the surface. In any of the above described process performed in a solid phase recation system which comprises a surface, the surface may be planar or may be a microbead, preferably 5 the surface comprises silicon, glass, a gel-based material and / or a polymeric material such as polystyrene, more preferably the surface is a microbead comprising a polymeric material such as polystyrene. In any such process, the surface may be bound to a substrate, preferably the surface is planar and / or the substrate comprises glass. The surface, e.g. a planar surface and / or the substrate may be provided as a flow-cell. 10          The invention provides a processes for performing a biochemical reaction within a cell in culture by introducing at least one of any of the above-described IDR-macromolecules or at least one of any of the above-described IDR-polypeptides into a cultured host cell, or by expressing at least one of any of the above-described IDR-polypeptides in the cultured host cell, to increase the efficiency of the biochemical 15   reaction within the cultured host cell. Any of the above-described processes for performing an in vitro biochemical reaction may comprise a biochemical reaction performed within a cell in culture, such as by introducing the at least one IDR-macromolecule or the at least one IDR-polypeptide into a cultured host cell, or by expressing the at least one IDR-polypeptide 20 in the cultured host cell, to increase the efficiency of the biochemical reaction within the cultured host cell. The biochemical reaction may be any reaction which leads to the manipulation of a nucleic acid molecule within the cultured host cell, or which leads to the alteration of a nucleic acid molecule within the cultured host cell, such as a change in the structure 25   of a nucleic acid molecule, such as a change in the nucleotide sequence of a nucleic acid molecule. The biochemical reaction may be any reaction which leads to the synthesis of a nucleic acid molecule in the cultured host cell. The biochemical reaction may be any reaction which leads to the expression of a polypeptide from a nucleic acid molecule. The biochemical reaction may be any reaction which leads to the editing of a nucleic 30 acid sequence within the cultured host cell, e.g. wherein the IDR-polypeptide is a CRISPR polypeptide, such as a Cas polypeptide, including a Cas9 polypeptide). The 2026234101   18 Sep 2026 biochemical reaction may be any reaction which leads to the cleavage of a nucleic acid within the cultured host cell. The biochemical reaction may be any reaction which leads to the homologous recombination of nucleic acids within the cultured host cell. The biochemical reaction may be a metabolic reaction within the cultured host cell to 5 produce one or more biological products of interest within the cultured host cell, or to produce one or more biological products of interest which are secreted from the cultured host cell or otherwise released from the cultured host cell into the culture media. In any one of the above-described processes, increasing the efficiency of the biochemical reaction may comprise increasing the efficiency of the reaction using the at 10   least one IDR-macromolecule or the at least one IDR-polypeptide compared to the efficiency of the reaction obtained by performing the reaction under the same conditions but wherein the relevant at least one macromolecule or the at least one polypeptide does not comprise or has not been tagged with one or more functional intrinsically disordered region polypeptide sequences, optionally wherein the reaction is performed in the 15 absence of an exogenously added crowding agent. In any one of the above-described RPA processes, increasing or enhancing the efficiency or performance of an RPA biochemical reaction may comprise increasing the amount of amplified product obtained in the RPA reaction using the at least one IDR-polypeptide compared to the amount of amplified product obtained by performing the 20 reaction under the same conditions but wherein the relevant at least one polypeptide has not been tagged with one or more functional intrinsically disordered region polypeptide sequences, optionally wherein the reaction is performed in the absence of an exogenously added crowding agent. In any one of the above-described processes involving introducing at least one 25 IDR-macromolecule or IDR-polypeptide into the in vitro reaction system, the efficiency of the reaction in the system is increased by the IDR-macromolecule or the IDR-polypeptide compared to the efficiency of the reaction in the system following the introduction of the at least one macromolecule or polypeptide under the same reaction conditions except that the at least one macromolecule or polypeptide does not comprise 30 one or more functional intrinsically disordered regions (IDRs). 2026234101   18 Sep 2026 In any one of the above-described processes involving introducing at least at least one polypeptide tagged with an amino acid sequence comprising or consisting of one or more functional intrinsically disordered regions (IDRs) (IDR-polypeptide) into the in vitro reaction system the efficiency of the reaction in the system is increased by 5 the IDR-polypeptide compared to the efficiency of the reaction in the system following the introduction of the at least one polypeptide under the same reaction conditions except that the at least one polypeptide has not been tagged with the amino acid sequence comprising or consisting of the one or more functional IDRs. The invention also provides a non-naturally occurring IDR-macromolecule 10   comprising a macromolecule and a tag amino acid sequence, wherein the tag amino acid sequence comprises or consists of one or more functional intrinsically disordered regions (IDRs), wherein the IDR-macromolecule is capable of causing liquid-liquid demixing in an aqueous in vitro reaction system. Any such IDR-macromolecule may be capable of causing liquid-liquid demixing and the formation of a plurality of phase-15 separated aqueous compartments in the system, preferably a plurality of detectable phase-separated aqueous particles. Any such liquid-liquid demixing caused by any such non-naturally occurring IDR-macromolecule in the in vitro reaction system may thereby increase the efficiency of the biochemical reaction. Any one of the above-described IDR-macromolecules may be a non-naturally 20 occurring, artificial or genetically engineered IDR-macromolecule or IDR-polypeptide comprising a macromolecule or polypeptide and the tag amino acid sequence. In the case of an IDR-polypeptide, the tag amino acid sequence may be located at the C-terminus of the polypeptide, at the N-terminus of the polypeptide or at both the C-terminus of the polypeptide and the N-terminus of the polypeptide, or at any amino acid 25 position along the length of the polypeptide. In any one of the above-described IDR-macromolecules or IDR-polypeptides, the one or more functional IDRs of the tag amino acid sequence are functional IDRs as defined in any one of the above-described processes. In any one of the above-described IDR-macromolecules or IDR-polypeptides the 30 tag sequence comprises amino acid residues which are capable of engaging in aromatic cation-pi interactions with multivalent metal cations, preferably divalent metal cations, 2026234101   18 Sep 2026 more preferably Mg2+, Mn2+, Ca2+, Co2+ or Ni2+ ions, yet more preferably Mg2+, Mn2+ or Ca2+, even more preferably Mg2+. In any one of the above-described IDR-macromolecules or IDR-polypeptides the IDR-macromolecule or the IDR-polypeptide comprises or consists of the 5 macromolecule or polypeptide tagged with an amino acid sequence which comprises or consists of the amino acid sequence of any one of SEQ ID NOs 1 to 43, or which comprises or consists of a functional variant amino acid sequence of SEQ ID NOs 1 to 43, e.g. which has 80% or more identity to any one of SEQ ID NOs 1 to 43. In any one of the above-described IDR-polypeptides, the polypeptide to which 10 the sequence comprising or consisting of the one or more functional IDRs is tagged may be an enzyme, such as a helicase, a gyrase, a recombinase, e.g. an RPA recombinase agent, a nuclease, e.g. an exonuclease and an endonuclease, a ligase, a glycolyase, a methylase, a methyltransferase, a glucosyltransferase, a polymerase, a kinase, a phosphatase, a gene editing enzyme such as a CRISPR enzyme, e.g. a Cas9 enzyme; a 15 co-factor, e.g. as an RPA recombinase loading protein and an RPA single strand stabilizing agent. The polypeptide to which the sequence comprising or consisting of the one or more functional IDRs is tagged may be a ligase, optionally an RB69 ligase, such as RB69 Iigase-His2 (SEQ ID NO:112). The polypeptide to which the sequence comprising or consisting of the one or more functional IDRs is tagged may be an RPA 20 single strand stabilizing agent, preferably Gp32; optionally wherein the IDR-polypeptide has the amino acid sequence of any one of SEQ ID NOs 65 to 88 and SEQ ID NO: 120, or wherein the IDR-polypeptide is a functional variant thereof, e.g. an IDR-polypeptide having an amino acid sequence which has 80% or more identity to any one of SEQ ID NOs 65 to 88 and SEQ ID NO: 120. The polypeptide to which the sequence 25 comprising or consisting of the one or more functional IDRs is tagged may be an RPA recombinase agent, preferably UvsX; optionally wherein the IDR-polypeptide has the amino acid sequence of any one of SEQ ID NOs 44 to 59, or wherein the IDR-polypeptide is a functional variant thereof, e.g. an IDR-polypeptide having an amino acid sequence which has 80% or more identity to any one of SEQ ID NOs 44 to 59. 30 The polypeptide to which the sequence comprising or consisting of the one or more functional IDRs is tagged may be an RPA recombinase loading protein, preferably 2026234101   18 Sep 2026 UvsY; optionally wherein the IDR-polypeptide has the amino acid sequence of any one of SEQ ID NOs 60 to 64, or wherein the IDR-polypeptide is a functional variant thereof, e.g. an IDR-polypeptide having an amino acid sequence which has 80% or more identity to any one of SEQ ID NOs 60 to 64. 5          The invention also provides an isolated nucleic acid molecule comprising a first nucleic acid sequence encoding any of the above-described IDR-polypeptides; optionally including a second nucleic acid sequence encoding a promoter, wherein the first nucleic acid sequence is operably linked to the second nucleic acid sequence. The invention also provides a recombinant polynucleotide expression vector comprising any 10 such nucleic acid molecule. The invention also provides a host cell comprising any such nucleic acid molecule, or any such recombinant polynucleotide expression vector. The invention also provides a cell culture comprising a growth medium and a population of any such host cells. The invention also provides a kit comprising any of the above-described non-15 naturally occurring IDR-macromolecules or IDR-polypeptides. Any such kit may further comprising additional RPA components comprising an RPA recombinase agent, and / or an RPA recombinase loading protein, and / or polymerase, and / or first and second nucleic acid primers, and / or an exonuclease, and / or a buffer, and / or a source of multivalent metal ions, preferably divalent metal cations. In any such kit, all 20 components may be provided in lyophilized form. The invention also provides a process of stimulating or enhancing liquid-liquid demixing in a solution, the process comprising providing a solution comprising any of the above-described IDR-macromolecules or any of the above-described IDR-polypeptides, and contacting the IDR-macromolecule or IDR-polypeptide in solution 25 with multivalent metal ions whereupon liquid-liquid demixing in the solution is stimulated or enhanced. The invention also provides a further process of stimulating or enhancing in an aqueous in vitro reaction system liquid-liquid demixing caused by an IDR-macromolecule or an IDR-polypeptide, the process comprising providing any one of the above-described IDR-macromolecules or any one of the above-described IDR-30 polypeptides into the system, providing multivalent metal ions into the system and allowing the IDR-macromolecule or IDR-polypeptide to contact the multivalent metal 2026234101   18 Sep 2026 ions whereupon liquid-liquid demixing caused by the IDR-macromolecule or the IDR-polypeptide in the solution is stimulated or enhanced. In any such processes the liquidliquid demixing may result in the formation of a plurality of phase-separated aqueous compartments, preferably a plurality of detectable phase-separated aqueous particles in 5 the solution. In any such process the multivalent metal ions may be divalent metal ions, optionally Mg2+, Mn2+, Ca2+, Co2+ or Ni2+, preferably Mg2+, Mn2+ or Ca2+, more preferably Mg2+. In any such further process, the multivalent metal ions may engage in aromatic cation-pi interactions with amino acid residues in the one or more functional IDR amino acid sequences, thereby promoting liquid-liquid demixing. 10           In any such further process, the conditions suitable for performing the reaction may further comprise providing ATP to the IDR-macromolecule or the 1DR-polypeptide in the in vitro reaction system, thereby further simulating or enhancing the liquid-liquid demixing and the formation of the plurality of phase-separated aqueous compartments caused by the IDR-macromolecule or the IDR-polypeptide and thereby 15   further increasing the efficiency of the biochemical reaction in the system, wherein ATP is provided in the system at a concentration of ImM to 3.5mM, optionally ImM to 2mM, preferably ImM. In any such further process, the conditions suitable for performing the reaction may further comprise providing multivalent metal ions to the IDR-polypeptide thereby 20 further stimulating or enhancing molecules necessary for the performance of the reaction to co-localise with the IDR-macromolecule or the IDR-polypeptide within the plurality of phase-separated aqueous compartments thereby further increasing the efficiency of the biochemical reaction in the system; optionally wherein the multivalent metal ions are provided at a concentration of about 22 mM or more, preferably wherein 25 the multivalent metal ions are provided at a concentration of between about 22 mM to 50 mM. The multivalent metal ions maybe divalent metal ions, optionally Mg2+, Mn2+, Ca2+, Co2+ or Ni2+, preferably Mg2+, Mn2+ or Ca2+, more preferably Mg2+. In any such further process, the conditions suitable for performing the reaction may further comprise providing ATP to the IDR-macromolecule or IDR-polypeptide in 30 the in vitro reaction system, thereby further stimulating or enhancing molecules necessary for the performance of the reaction to co-localise with the IDR-polypeptide 2026234101   18 Sep 2026 within the plurality of phase-separated aqueous compartments and thereby further increasing the efficiency of the biochemical reaction in the system, wherein ATP is provided in the system at a concentration of ImM to 3.5mM, optionally ImM to 2mM, preferably ImM. 5          The invention also provides the use of multivalent metal ions in stimulating or enhancing liquid-liquid demixing in a solution, said demixing being mediated by any one of the above-described IDR-macromolecules or any one of the above-described IDR-polypeptides. The invention also provides the use of multivalent metal ions in stimulating or enhancing in an aqueous in vitro reaction system liquid-liquid demixing 10   caused by an IDR-macromolecule or an IDR-polypeptide which has been introduced into the system, wherein said IDR-macromolecule or IDR-polypeptide is any one of the above-described IDR-macromolecules or any one of the above-described IDR-polypeptides. In any such use, said liquid-liquid demixing may result in the formation of a plurality of phase-separated aqueous compartments caused by the IDR- 15 macromolecule or IDR-polypeptide, preferably a plurality of detectable phase-separated aqueous particles in the solution. In any such use, the multivalent metal ions may be divalent metal ions, optionally Mg2+, Mn2+, Ca2+, Co2+ or Ni2+, preferably Mg2+, Mn2+ or Ca2+, more preferably Mg2+. In any such use, the multivalent metal ions may engage in aromatic cation-pi interactions with amino acid residues in the one or more functional 20 IDR amino acid sequences, thereby promoting liquid-liquid demixing. In any such use, the conditions suitable for performing the reaction may further comprise providing ATP to the IDR-macromolecule or the IDR-polypeptide in the in vitro reaction system, thereby further simulating or enhancing the liquid-liquid demixing and the formation of the plurality of phase-separated aqueous compartments 25 caused by the IDR-macromolecule or the IDR-polypeptide and thereby further increasing the efficiency of the biochemical reaction in the system, wherein ATP is provided in the system at a concentration of ImM to 3.5mM, optionally ImM to 2mM, preferably ImM. In any such use, the conditions suitable for performing the reaction may further 30 comprise providing multivalent metal ions to the IDR-polypeptide thereby further stimulating or enhancing molecules necessary for the performance of the reaction to co- 2026234101   18 Sep 2026 localise with the IDR-macromolecule or the IDR-polypeptide within the plurality of phase-separated aqueous compartments thereby further increasing the efficiency of the biochemical reaction in the system; optionally wherein the multivalent metal ions are provided at a concentration of about 22 mM or more, preferably wherein the multivalent 5 metal ions are provided at a concentration of between about 22 mM to 50 mM. The multivalent metal ions may be divalent metal ions, optionally Mg2+, Mn2+, Ca2+, Co2+ or Ni2+, preferably Mg2+, Mn2+ or Ca2+, more preferably Mg2+. In any such use, the conditions suitable for performing the reaction may further comprise providing ATP to the IDR-macromolecule or IDR-polypeptide in the in vitro 10   reaction system, thereby further stimulating or enhancing molecules necessary for the performance of the reaction to co-localise with the IDR-polypeptide within the plurality of phase-separated aqueous compartments and thereby further increasing the efficiency of the biochemical reaction in the system, wherein ATP is provided in the system at a concentration of ImM to 3.5mM, optionally ImM to 2mM, preferably ImM. 15          The invention also provides any one of the above-described non-naturally occurring IDR-macromolecules or any one of the above-described IDR-polypeptides for use in therapy, for use as a medicament, for use as a pharmaceutical, for use in a diagnostic method or for use as a diagnostic agent. The invention also provides a process for making any one of the above- 20 described non-naturally occurring IDR-macromolecules or any one of the abovedescribed IDR-polypeptides, the process comprising providing a macromolecule, optionally a polypeptide, and tagging the macromolecule or polypeptide with one or more functional intrinsically disordered region polypeptide sequences. Said tagging may be performed by any means described and defined herein. Said one or more 25 functional intrinsically disordered region polypeptide sequences may be any of the same as described and defined herein. Said macromolecule or polypeptide may be any suitable macromolecule or polypeptide, including any macromolecule or polypeptide described and defined herein. Any one of the above-described IDR-macromolecules or IDR-polypeptides may 30 increase the efficiency of the biochemical reaction. Increasing the efficiency of the biochemical reaction may comprise increasing the efficiency of the reaction using the 2026234101   18 Sep 2026 IDR-macromolecule or the IDR-polypeptide compared to the efficiency of the reaction obtained by performing the reaction under the same conditions but wherein the relevant macromolecule or the relevant polypeptide does not comprise or has not been tagged with one or more functional intrinsically disordered region polypeptide sequences, 5 optionally wherein the reaction is performed in the absence of an exogenously added crowding agent. Any one of the above-described IDR-macromolecules or IDR-polypeptides may increase the efficiency of a biochemical reaction, wherein the reaction in a recombinase polymerase amplification (RPA) reaction. Increasing the efficiency or performance of 10 an RPA biochemical reaction may comprise increasing the amount of amplified product obtained in the RPA reaction using the IDR-polypeptide compared to the amount of amplified product obtained by performing the reaction under the same conditions but wherein the relevant polypeptide does not comprise or has not been tagged with one or more functional intrinsically disordered region polypeptide sequences, optionally 15   wherein the RPA reaction is performed in the absence of an exogenously added crowding agent. The invention also provides a method for determining the nucleotide sequence of one or more target polynucleotide molecules, said method comprising the steps of: (i) performing an above-described process to amplify the one or more target 20 polynucleotide molecules thereby obtaining a population comprising a plurality of copies of the one or more target polynucleotide molecules; and (ii) performing one or more nucleic acid sequencing reactions on the population comprising the plurality of copies of the target polynucleotide molecules, preferably wherein the method is performed in a solid phase reaction system 25 which comprises a surface. The invention also provides the use of any one of the above-described IDR-macromolecules or any one of the above-described IDR-polypeptides in a method for determining the nucleotide sequence of one or more target polynucleotide molecules, preferably wherein the method is as described above. 30          The invention also provides a polypeptide or an isolated polypeptide which comprises or consists of the amino acid sequence of any one of SEQ ID NOs 1 to 43, or 2026234101   18 Sep 2026 which comprises or consists of a functional variant amino acid sequence of any one of SEQ ID NOs 1 to 43, e.g. which has 80% or more identity to any one of SEQ ID NOs 1 to 43. Any such polypeptide can be attached to / tagged to a macromolecule or a polypeptide to form an IDR-tagged macromolecule or an IDR-tagged polypeptide as 5 described further herein. The macromolecule or polypeptide which is tagged may be a macromolecule or polypeptide required for the performance of a biochemical reaction in an aqueous reaction system. When maintained within the aqueous reaction system under conditions for performing the biochemcical reaction, any such IDR-tagged macromolecule or IDR-tagged polypeptide can cause liquid-liquid demixing caused by 10 the amino acid sequence of any one of SEQ ID NOs 1 to 43 or any functional variant thereof thereof and the formation of a plurality of phase-separated aqueous compartments, preferably a plurality of detectable phase-separated aqueous particles, in the system thereby increasing the efficiency of the biochemical reaction in the system. When maintained within the aqueous reaction system under conditions for performing 15   the biochemcical reaction, any such IDR-tagged macromolecule or IDR-tagged polypeptide causes molecules necessary for the performance of the reaction to colocalise with the IDR-macromolecule or the IDR-polypeptide within the plurality of phase-separated aqueous compartments, thereby increasing the efficiency of the biochemical reaction in the system. 20          The aqueous reaction system may be an aqueous in vitro reaction system. The invention also provides an isolated nucleic acid molecule comprising a nucleic acid sequence encoding a polypeptide which comprises or consists of the amino acid sequence of any one of SEQ ID NOs 1 to 43, or which comprises or consists of a functional variant amino acid sequence of SEQ ID NOs 1 to 43, e.g. which has 80% or 25 more identity to any one of SEQ ID NOs 1 to 43. The invention also provides the use of an IDR moiety, which is a polypeptide which comprises or consists of one or more functional intrinsically disordered regions (IDRs), in producing an IDR-tagged macromolecule or IDR-tagged polypeptide wherein the IDR moiety is attached to / tagged to a macromolecule or a polypeptide; 30 wherein the macromolecule or polypeptide which is tagged is a macromolecule or polypeptide required for the performance of a biochemical reaction in an aqueous 2026234101   18 Sep 2026 reaction system, and wherein when maintained within the aqueous reaction system under conditions for performing the biochemcical reaction, the IDR-tagged macromolecule or IDR-tagged polypeptide causes liquid-liquid demixing caused by the IDR moiety and causes the formation of a plurality of phase-separated aqueous 5 compartments, preferably a plurality of detectable phase-separated aqueous particles, in the system thereby increasing the efficiency of the biochemical reaction in the system. When maintained within the aqueous reaction system under conditions for performing the biochemcical reaction, any such IDR-tagged macromolecule or IDR-tagged polypeptide causes molecules necessary for the performance of the reaction to co- 10   localise with the IDR-tagged macromolecule or the IDR-tagged polypeptide within the plurality of phase-separated aqueous compartments, thereby increasing the efficiency of the biochemical reaction in the system. Preferably, the IDR moiety is attached to / tagged to a polypeptide, thereby producing an IDR-tagged polypeptide, preferably produced as a recombinant genetic 15    fusion protein. Preferably, the IDR moiety is a polypeptide which comprises or consists of the amino acid sequence of any one of SEQ ID NOs 1 to 43, or a polypeptide which comprises or consists of a functional variant amino acid sequence of any one of SEQ ID NOs 1 to 43, e.g. which has 80% or more identity to any one of SEQ ID NOs 1 to 43. 20         Any of the above-described IDR-tagged macromolecules or IDR-tagged polypeptides may be defined as non-naturally occurring, artificial or genetically engineered macromolecules or polypeptides. Any of the above-described IDR-tagged macromolecules or IDR-tagged polypeptides may further possess the characteristic features of any one or more of the 25 IDR-macromolecules or IDR-polypeptides described and defined herein. The aqueous reaction system may be an aqueous in vitro reaction system. The invention further provides an IDR-tagged macromolecule or an IDR-tagged polypeptide obtained in accordance with any of the above-described uses. The invention also provides a method for producing an IDR-tagged 30 macromolecule or an IDR-tagged polypeptide comprising providing a macromolecule or a polypeptide and attaching / tagging an IDR moiety thereto, wherein the IDR moiety 2026234101   18 Sep 2026 is a polypeptide which comprises or consists of one or more functional intrinsically disordered regions (IDRs); wherein the macromolecule or polypeptide which is tagged is a macromolecule or polypeptide required for the performance of a biochemical reaction in an aqueous reaction system, and wherein when maintained within the 5 aqueous reaction system under conditions for performing the biochemcical reaction, the IDR-tagged macromolecule or IDR-tagged polypeptide causes liquid-liquid demixing caused by the IDR moiety and causes the formation of a plurality of phase-separated aqueous compartments, preferably a plurality of detectable phase-separated aqueous particles, in the system thereby increasing the efficiency of the biochemical reaction in 10 the system. When maintained within the aqueous reaction system under conditions for performing the biochemcical reaction, any such IDR-tagged macromolecule or IDR-tagged polypeptide causes molecules necessary for the performance of the reaction to co-localise with the IDR-tagged macromolecule or the IDR-tagged polypeptide within the plurality of phase-separated aqueous compartments, thereby increasing the 15 efficiency of the biochemical reaction in the system. Preferably, the method comprises providing a polypeptide and attaching / tagging an IDR moiety thereto to produce an IDR-tagged polypeptide, preferably produced as a recombinant genetic fusion protein. Preferably, the IDR moiety is a polypeptide which comprises or consists of the 20 amino acid sequence of any one of SEQ ID NOs 1 to 43, or a polypeptide which comprises or consists of a functional variant amino acid sequence of any one of SEQ ID NOs 1 to 43, e.g. which has 80% or more identity to any one of SEQ ID NOs 1 to 43. Any of the above-described IDR-tagged macromolecules or IDR-tagged polypeptides may be defined as non-naturally occurring, artificial or genetically 25 engineered macromolecules or polypeptides. Any of the above-described IDR-tagged macromolecules or IDR-tagged polypeptides may further possess the characteristic features of any one or more of the IDR-macromolecules or IDR-polypeptides described and defined herein. The aqueous reaction system may be an aqueous in vitro reaction system. 30          The invention further provides an IDR-tagged macromolecule or an IDR-tagged polypeptide obtained by any of the above-described methods. 2026234101   18 Sep 2026 BRIEF DESCRIPTION OF THE FIGURES Figure 1 shows real-time recombinase polymerase amplification traces using an IDR-tagged Gp32 fusion protein (Gp32-HIS2) at varying template nucleic acid 5 concentrations. Figure 2 shows real-time recombinase polymerase amplification traces using an IDR-tagged Gp32 fusion protein (Gp32-HIS5) at varying template nucleic acid concentrations. Figure 3 shows real-time recombinase polymerase amplification traces using an 10 IDR-tagged Gp32 fusion protein (Gp32-HRP1) at varying template nucleic acid concentrations. Figure 4 shows real-time recombinase polymerase amplification traces using an IDR-tagged Gp32 fusion protein (Gp32-Supl) at varying template nucleic acid concentrations. 15          Figure 5 shows real-time recombinase polymerase amplification traces using an IDR-tagged Gp32 fusion protein (Gp32-Sup2) at varying template nucleic acid concentrations. The experiments shown in Figures 5A, B, C and D use respectively a Gp32 fusion protein with one, two, three and four Sup2 IDR repeats. Figure 6 shows real-time recombinase polymerase amplification traces using an 20 IDR-tagged Gp32 fusion protein (Gp32-HIS5) at varying MgOAc concentrations. Figure 7 shows real-time recombinase polymerase amplification traces using an IDR-tagged Gp32 fusion protein (Gp32-HIS2) at varying phosphocreatine concentrations. Figure 8 shows real-time recombinase polymerase amplification traces using an 25 IDR-tagged Gp32 fusion protein (Gp32-HRP1) at varying KOAc concentrations. Figure 9 shows real-time recombinase polymerase amplification traces using Gp32 tagged with seven histidine residues (for protein purification purpose, i.e. no IDR tag) compared with an IDR-tagged Gp32 fusion protein (Gp32-Supl) either in the presence or absence of a crowding agent (PEG). 30          Figure 10 shows the effect of multivalent metal cations on the promotion of phase separation (particle formation) mediated by IDR amino acid sequence tags in the 2026234101   18 Sep 2026 absence of a crowding agent. The IDR amino acid sequences were tagged to the Gp32 protein to create the Gp32-HIS2 fusion protein (Figure 10A), the Gp32-HRP1 fusion protein (Figure 10B), the Gp32-Supl fusion protein (Figure 10C) and the Gp32-Fib fusion protein (Figure 10D). In each case the effect of representative concentrations of 5 divalent metal cations, i.e. magnesium (MgOAc), manganese (MgCh) and calcium (CaCh) was tested. Figure 11 shows the effect of multivalent metal cations on the promotion of phase separation (particle formation) mediated by IDR amino acid sequence tags in the absence of a crowding agent. The IDR amino acid sequences were tagged to the Gp32 10 protein to create the Gp32-Fib fusion protein (Figure 11A), the Gp32-Supl fusion protein (Figure 1 IB), the Gp32-HIS2 fusion protein (Figure 1 IC), the Gp32-HRP1 fusion protein (Figure 1 ID), the Gp32-HIS5 fusion protein (Figure 1 IE). In each case the effect of representative concentrations of divalent metal cations, i.e. magnesium (MgOAc), manganese (MgCh) and calcium (CaCh) was tested. 15          Figure 12 shows the effect of a divalent metal cation, i.e. magnesium (MgOAc), on the capability of an IDR-tagged Gp32 fusion protein (Gp32-HRP1) to promote phase separation (particle formation) in an exemplary in vitro biochemical reaction environment in the absence of a crowding agent. Figure 13 shows the effect of a divalent metal cation, i.e. magnesium (MgOAc), 20 on the capability of an IDR-tagged Gp32 fusion protein (Gp32-HIS2) to promote phase separation (particle formation) in an exemplary in vitro biochemical reaction environment in the absence of a crowding agent. Figure 14 shows the effect of varying concentrations of a divalent metal cation, i.e. magnesium (MgOAc), on the capability of an IDR-tagged Gp32 fusion protein 25   (Gp32-HRP1) to promote phase separation (particle formation) in an exemplary in vitro biochemical reaction environment in the absence of a crowding agent. Figure 15 shows the effect of varying concentrations of a divalent metal cation, i.e. magnesium (MgOAc), on the capability of an IDR-tagged Gp32 fusion protein (Gp32-HIS2) to promote phase separation (particle formation) in an exemplary in vitro 30 biochemical reaction environment in the absence of a crowding agent. 2026234101   18 Sep 2026 Figure 16 shows the effect of addition of a divalent metal cation, i.e. magnesium (MgOAc), on the capability of an IDR-tagged Gp32 fusion protein (Gp32-HRP1) to promote phase separation in an exemplary in vitro biochemical reaction environment in the absence of a crowding agent. Phase separation is demonstrated by 5 the formation of an opaque solution following the addition of MgOAc due to particle formation (Figure 16A) and particle formation is further demonstrated by pelleting of the particles (Figure 16B). RPA protein components are demonstrated to associate with particles as revealed by SDS-PAGE analysis of pelleted material. Figure 17 shows real-time recombinase polymerase amplification traces using a 10 native Gp32 fusion protein either in the presence of absence of crowding agent. The experiment reveals that Gp32 which is not tagged with an amino acid sequence comprising an intrinsically disordered region (IDR) is unable to mediate amplification in the absence of a crowding agent. Figure 18. Figure 18A is a cartoon depicting a reaction mixture set up for real-15 time amplification using dual-primer beads. Figure 18B is a cartoon depicting amplified products in the real-time reaction. Figure 18C is a cartoon depicting amplification characterisation in the end-point reaction. Figure 18D shows real-time recombinase polymerase amplification traces using an IDR-tagged Gp32 fusion protein (Gp32-HIS2) using primers attached to a solid surface or using primers free in solution. 20 Figures 18E and F show end-point recombinase polymerase amplification traces using an IDR-tagged Gp32 fusion protein (Gp32-HIS2) using primers attached to a solid surface. Figure 19 shows disorder profiles generated using the MetaDisorder software program for Gp32 (Figure 19A), UvsY (Figure 19B) and UvsX (Figure 19C). 25          Figure 20 shows the effect of varying concentrations of a divalent metal cation, i.e. magnesium (MgCh), on the capability of an IDR-tagged RB69 ligase fusion protein (RB69 ligase-HIS2) to promote phase separation (particle formation) in an exemplary in vitro biochemical reaction environment in the absence of a crowding agent. Figure 21 shows the ligase activity performance of an IDR-tagged RB69 ligase 30 fusion protein (RB69 ligase-HIS2) in an exemplary in vitro biochemical reaction environment in the absence of a crowding agent. 2026234101   18 Sep 2026 Figure 22 shows the ligase activity performance of an IDR-tagged RB69 ligase fusion protein (RB69 ligase-HIS2) in an exemplary in vitro biochemical reaction environment in the absence of a crowding agent compared with untagged RB69 ligase and T4 DNA ligase. 5          Figure 23 shows the ligase activity performance of an IDR-tagged RB69 ligase fusion protein (RB69 ligase-HIS2) in an exemplary in vitro biochemical reaction environment in the absence of a crowding agent compared with NEBNext Ultra II ligation master mix. Figure 24 shows the effect of ATP on the capability of an IDR-tagged RB69 10 ligase fusion protein (RB69 ligase-HIS2) to promote phase separation (particle formation) in an exemplary in vitro biochemical reaction environment in the absence of a crowding agent. Figure 25 shows brightfield and fluorescent images of representative sections of FlexWell™ chambers that had 0, 5, 10, 20, 40 or 80 copies of single stranded UP1- 15 UP2'-TF1L template per bead annealed at 50°C for 1 hour and then amplified by recombinase polymerase amplification using an IDR-tagged Gp32 fusion protein (Gp32-Hrpl) using primers attached to a solid surface in the absence of a crowding agent such as PEG. Amplification was detected by nicking amplicons with Nt. BbvCI and extending the nick with Aminoallyl-dUTP-XX-ATTO-594. No fluorescence was 20 observed on beads where template had not been added and an increasing amount of fluorescence was observed on beads where increasing amounts of template had been annealed. This shows that amplification occurred on the solid surface of the beads in the absence of a crowding agent. Figure 26 A shows brightfield and fluorescent images demonstrating the 25 formation of phase-separated aqueous particles mediated by an IDR-tagged Gp32 fusion protein (Gp32-Hrpl). Figure 26 B shows a plot demonstrating increased efficiency of a reaction (rate of nucleic acid cutting by Cas 12a) upon the formation of phase-separated aqueous particles mediated by Gp32-Hrpl. 30 2026234101   18 Sep 2026 DETAILED DESCRIPTION OF THE INVENTION Recombinase polymerase amplification is a technique for amplifying nucleic acid molecules. The system utilises, inter alia, a recombinase enzyme and preferably a 5 recombinase loading protein. These protein components form a complex with amplification primers. Following binding to the target nucleic acid molecule to be amplified, the complex “scans” the target nucleic acid molecule and “searches” for regions of complementarity between the target and primer sequences. Once a complementary region is found the complex facilitates the binding of the primer to the 10 target sequence. A polymerase enzyme can then extend the primer to generate a copy of the target sequence. The use of the recombinase complex provides a key difference over other nucleic acid amplification methods such as PCR. In RPA there is no requirement for melting and annealing steps driven by thermal cycling, as the recombinase complex provides an entirely enzymatic-based solution to the issue of 15   primer binding. RPA is thus an isothermal technique. The absence of a requirement for extreme thermal cycling means that RPA has many obvious advantages over techniques such as PCR. A well-documented requirement in RPA methods is the presence of a “crowding agent”, also commonly referred to in the technical field as a “macromolecular crowding 20 agent”. These agents are well known in the technical field and have an art-understood meaning. Crowding agents are discussed in more detail herein. One of the most commonly used crowding agents in RPA methods is polyethylene glycol (PEG), although other crowding agents can be used. Prior to the present invention the use of a crowding agent was considered to be an essential requirement in RPA methods. 25           The present inventors have surprisingly discovered that it is possible to bypass the hitherto-perceived critical requirement for a crowding agent in RPA methods. The present invention is founded on this discovery. The inventors have surprisingly discovered that by “tagging” a macromolecule, such as a protein component required in RPA methods, with amino acid sequences 30 which comprise or consist of one or more functional “intrincically disordered regions” (IDRs), the IDR amino acid sequence tag is capable of facilitating efficient RPA in the 2026234101   18 Sep 2026 complete absence of a crowding agent. As such, efficient amplification can be achieved in an RPA system without reliance on a crowding agent, thus reducing the complexity of RPA reactions. The inventors have also surprisingly discovered that the efficiency of 5 amplification in RPA methods involving an IDR-tagged macromolecular component in the absence of a crowding agent can be correlated with the functional capability of the IDR amino acid tag sequence to promote liquid-liquid demixing leading to phase separation in the biochemical reaction system. Phase separation may be assessed by the formation in the biochemical reaction milieu of phase-separated aqueous compartments, 10 particularly spherical-like aqueous globular foci or phase-separated aqueous particles which are amenable to detection by standard methods, including by microscopic observation, as described further herein. Furthermore, the inventors have also surprisingly discovered that the provision of an IDR-tagged macromolecular component and a crowding agent can provide 15   additive and even synergistic effects with respect to the efficiency of amplification in RPA methods. Yet further, the inventors have surprisingly discovered that the efficiency of amplification in RPA methods involving an IDR amino acid-tagged macromolecular component in the absence of a crowding agent can be correlated with the concentration 20 of multivalent metal cations introduced into the biochemical reaction milieu. Thus multivalent metal cations can further stimulate or enhance liquid-liquid demixing caused by IDR-macromolecules or IDR-polypeptides and thereby further increase reaction efficiency. The inventors have also surprisingly discovered that certain concentrations of 25 ATP, as described further herein, can further stimulate or enhance liquid-liquid demixing caused by IDR-macromolecules or IDR-polypeptides and thereby further increase reaction efficiency. Based on these surprising discoveries the invention provides methods and reagents which increase the efficiency of enzymatic-based in vitro biochemical 30 reactions, including RPA reactions, as further described herein. 2026234101   18 Sep 2026 The IDR amino acid sequences and IDR reagents described and defined herein have broad applicability as useful reagents to be applied to any suitable macromolecular component of a biochemical reaction, such as a polypeptide, thereby promoting liquidliquid demixing and incuding promoting phase separation in the biochemical reaction 5 milieu without reliance on macromolecular crowding agents, particularly when the IDR amino acid sequences are used in concert with a multivalent metal cation. The invention further embraces the use of a multivalent metal cation, such as a divalent metal cation, or any functional equivalent thereof, in promoting IDR amino acid sequence-mediated phase separation in the biochemical reaction milieu, without 10 reliance on macromolecular crowding agents. The invention thus provides IDR-based processes, macromolecules, polypeptides, nucleic acids, vectors, host cells and uses, as described and defined furthere herein. Elements of the invention are described in turn below. 15 Biochemical reactions As explained above, the inventors have surprisingly discovered that it is possible to bypass the requirement for a crowding agent, a previously-considered essential 20 component of RPA and other reactions. As described in detail herein, this may be achieved by attaching / tethering / tagging an amino acid sequence comprising one or more functional intrinsically disordered regions (IDRs) to a protein component required in the RPA reaction. The inventors have also surprisingly shown that a functional intrinsically disordered region attached to a ligase enzyme is capable of increasing the 25 efficiciency of a ligase reaction. The inventors have shown that the degree of phase separation induced by the IDR amino acid sequence can be correlated with the efficiency of the reaction, e.g. amplification, in the absence of a crowding agent, and can be enhanced with a multivalent metal cation. Based on these surprising observations it is plausibly expected that such IDR amino acid sequences associated 30 with a macromolecule or protein component of a biochemical reaction will improve the 2026234101   18 Sep 2026 efficiency of the reaction in the in vitro or in vivo biochemical reaction environment, particularly in the absence of an added / exogenous crowding agent. Accordingly, the invention embraces the use of any of the IDR amino acid sequences described and defined herein to be applied to any suitable macromolecular or 5 polypeptide component of an in vitro or in vivo biochemical reaction, thus providing IDR reagents capable of promoting liquid-liquid demixing in the biochemical reaction milieu and increasing the efficiency of the biochemical reaction. Such liquid-liquid demixing in the biochemical reaction milieu may result in phase separation the biochemical reaction milieu. Such liquid-liquid demixing in the biochemical reaction 10 milieu may result in phase separation leading to, causing or promoting the formation of phase-separated aqueous compartments, including detectable phase-separated aqueous particles in the biochemical reaction milieu, as described further herein. Such IDR reagents, or IDR-based reagents, as described and defined further herein may be referred to interchangeably to describe any one or more of an IDR-macromolecule or 15 IDR-tagged macromolecule, or an IDR-polypeptide or IDR-tagged polypeptide. In any one of the methods, processes and uses, or in any one of the non-naturally occurring IDR-macromolecules, IDR-fusion macromolecules or isolated nucleic acid molecules encoding the same, recombinant polynucleotide expression vectors or host cells described and defined herein, increasing or enhancing the efficiency or 20 performance of a biochemical reaction may comprise increasing the efficiency of the reaction using any one or more of the IDR-based macromolecules or polypeptides described herein compared to the efficiency obtained by performing the reaction under the same conditions but wherein the relevant macromolecule or polypeptide does not comprise or has not been tagged with one or more functional intrinsically disordered 25 region polypeptide sequences, optionally wherein the reaction is performed in the absence of an exogenously added crowding agent. Increasing or enhancing the efficiency or performance of a biochemical reaction is to be understood according to generally accepted concepts. For example, reaction efficiency in an RPA reaction, or in any other nucleic acid amplification reaction, may 30 be undersood as providing an equivalent total population of amplicons using comparatively less starting target nucleic acid, or providing a comparatively faster time 2026234101   18 Sep 2026 to detection or a comparatively faster rate of amplification using the same amount of starting target nucleic acid. Increasing or enhancing the efficiency or performance of an RPA biochemical reaction may comprise increasing the amount of amplified product obtained in the RPA 5 reaction using any one or more of the IDR-based macromolecules or polypeptides described herein compared to the amount of amplified product obtained by performing the reaction under the same conditions but wherein the relevant macromolecule or polypeptide does not comprise or has not been tagged with one or more functional intrinsically disordered region polypeptide sequences, and optionally wherein the 10 reaction is performed in the absence of an exogenously added crowding agent. Increasing the efficiency of a biochemical reaction in a reaction system, such as an in vitro reaction system, may comprise increasing any measurable parameter of the reaction in the system over a specified time period, such as the rate of the reaction over a time period, the amount of a substrate consumed over a time period, the amount of a 15   product produced over a time period etc. Increasing the efficiency of a biochemical reaction in a reaction system, such as an in vitro reaction system, may comprise increasing the parameter of the reaction within detectable phase-separated compartments, such as detectable phase-separated aqueous particles. This can, for example, be inferred indirectly by measuring the 20 parameter of the reaction and correlating the increase with formation of detectable phase-separated aqueous particles and / or the detectable co-localisation of reaction molecules into detectable phase-separated aqueous particles. Described herein are straightforward bioinformatics methods and phase separation assays which can be used to establish whether any IDR amino acid sequence 25 is capable of functioning in the required manner of promoting liquid-liquid demixing and phase separation in the desired biochemical reaction milieu when used with a given a macromolecule or protein and included in the desired in vitro biochemical reaction environment. Moreover the suitability of any given co-factor, in particular a multivalent, e.g. divalent, metal cation, may be established in these assays in a very 30 straightforward way. 2026234101   18 Sep 2026 Accordingly, the invention provides IDR reagents as described and defined herein that may usefully be applied in any given desired in vitro or in vivo biochemical reaction environment. Any of the IDR amino acid sequences as described and defined herein may be 5 used with any macromolecular or protein component required for the performance of an in vitro or in vivo biochemical reaction, such as any of the reactions described herein. Any of the IDR amino acid sequences as described and defined herein may be used with any macromolecular or protein component required for the performance of a nucleic acid synthesis reaction. 10          Any of the IDR amino acid sequences as described and defined herein may be used with any macromolecular or protein component required for the performance of a nucleic acid synthesis reaction wherein a polymerase is used to synthesize a new nucleic acid molecule by extending a primer nucleic acid molecule. Any of the IDR amino acid sequences as described and defined herein may be 15   used with any macromolecular or protein component required for the performance of a nucleic acid amplification reaction. The nucleic acid amplification reaction may be a reaction which involves thermal cycling. The nucleic acid amplification reaction may be an isothermal amplification reaction. The nucleic acid amplification reaction may be polymerase chain reaction (PCR), polymerase spiral reaction (PSR), loop mediated 20 isothermal amplification (LAMP), nucleic acid sequence based amplification (NASBA), self-sustained sequence replication (3SR), rolling circle amplification (RCA), strand displacement amplification (SDA), multiple displacement amplification (MDA), ligase chain reaction (LCR), helicase dependant amplification (HDA), ramification amplification method (RAM), recombinase polymerase amplification (RPA), 25 transcription-mediated amplification (TMA) or nicking enzyme amplification reaction (NEAR). Any of the IDR amino acid sequences as described and defined herein may be used with any macromolecular or protein component required for the performance of a gene editing reaction. 2026234101   18 Sep 2026 Any of the IDR amino acid sequences as described and defined herein may be used with any macromolecular or protein component required for the performance of a CRISPR reaction. Any of the IDR amino acid sequences as described and defined herein may be 5 used with any macromolecular or protein component required for the performance of a prime editing gene editing reaction, wherein a CRISPR enzyme, such as a Cas enzyme, e.g. Cas9, is provided in a complex with at least a reverse transcriptase enzyme, optionally further with a prime editing guide RNA (pegRNA), and wherein any component of the prime editing complex may be provided tagged with one or more 10 functional intrinsically disordered region (IDR) polypeptide sequences, e.g. wherein the CRISPR enzyme is tagged with the one or more functional IDR polypeptide sequences or wherein the reverse transcriptase enzyme is tagged with the one or more functional IDR polypeptide sequences. Any of the IDR amino acid sequences as described and defined herein may be 15   used with any macromolecular or protein component required for the performance of a ligation reaction. Any of the IDR amino acid sequences as described and defined herein may be used with any macromolecular or protein component required for the performance of an exonuclease reaction. 20         Any of the IDR amino acid sequences as described and defined herein may be used with any macromolecular or protein component required for the performance of an endonuclease reaction, a transcription reaction, a DNA methylation reaction, a DNA glycosylation reaction, an antibody-antigen reaction, a drug-target reaction. Any of the IDR amino acid sequences as described and defined herein may be 25 used with any macromolecular or protein component required for the performance of a reaction involving protein:protein interactions. A process for performing an in vitro biochemical reaction as used herein is intended to embrace biochemical reactions performed directly in solution in a reaction vessel, such as an RPA reaction as described further herein. 30          A process for performing an in vitro biochemical reaction as used herein also includes biochemical reactions performed within cells in culture, such as by expressing 2026234101   18 Sep 2026 an IDR reagent as defined herein in a cultured host cell to increase the efficiency of a biochemical reaction within the cultured host cell. A process for performing an in vitro biochemical reaction as used herein includes a biochemical reaction performed within a host cell in culture by introducing 5 an IDR reagent as defined herein into a cultured host cell or expressing an IDR reagent as defined herein in a cultured host cell to increase the efficiency of a biochemical reaction within the cultured host cell, wherein the biochemical reaction is any reaction which leads to the manipulation of a nucleic acid molecule within the cultured host cell, or which leads to the alteration of a nucleic acid molecule within the cultured host cell, 10   such as a change in the structure of a nucleic acid molecule, such as a change in the nucleotide sequence of a nucleic acid molecule. A process for performing an in vitro biochemical reaction as used herein includes a biochemical reaction performed within cells in culture by introducing an IDR reagent as defined herein into a cultured host cell or expressing an IDR reagent as 15   defined herein in a cultured host cell to increase the efficiency of a biochemical reaction within the cultured host cell, wherein the biochemical reaction is any reaction which leads to the synthesis of a nucleic acid molecule in the cultured host cell. A process for performing an in vitro biochemical reaction as used herein includes biochemical reactions performed within cells in culture by introducing an IDR 20 reagent as defined herein into a cultured host cell or expressing an IDR reagent as defined herein in a cultured host cell to increase the efficiency of a biochemical reaction within the cultured host cell, wherein the biochemical reaction is any reaction which leads to the expression of a polypeptide from a nucleic acid molecule. A process for performing an in vitro biochemical reaction as used herein 25 includes biochemical reactions performed within cells in culture by introducing an IDR reagent as defined herein into a cultured host cell or expressing an IDR reagent as defined herein in a cultured host cell to increase the efficiency of a biochemical reaction within the cultured host cell, wherein the biochemical reaction is any reaction which leads to the editing of a nucleic acid sequence within the cultured host cell (e.g. wherein 30 the IDR-polypeptide is a CRISPR polypeptide, such as a Cas polypeptide, including a Cas9 polypeptide, or wherein the IDR-polypeptide is a polypeptide in complex with a 2026234101   18 Sep 2026 CRISPR polypeptide, such as wherein the IDR-polypeptide is a reverse transcriptase enzyme), the cleavage of a nucleic acid within the cultured host cell and homologous recombination of nucleic acids within the cultured host cell. A process for performing an in vitro biochemical reaction as used herein 5 includes biochemical reactions performed within cells in culture by introducing an IDR reagent as defined herein into a cultured host cell or expressing an IDR reagent as defined herein in a cultured host cell to increase the efficiency of a biochemical reaction within the cultured host cell, wherein the biochemical reaction is a metabolic reaction within the cultured host cell to produce one or more biological products of interest 10 within the cultured host cell, or to produce one or more biological products of interest which are secreted from the cultured host cell or otherwise released from the cultured host cell into the culture media. The invention is also intended to embrace biochemical reactions performed ex vivo, for example by expressing the IDR reagents defined herein in cells of e.g. a tissue 15   culture or any other suitable complex biological system developed external to the body. Accodingly, any reference to a process for performing a biochemical reaction in an aqueous in vitro reaction system as used herein using any of the IDR reagents as defined herein may alternatively be defined as a process for performing a biochemical reaction in an aqueous ex vivo reaction system using any of the IDR reagents as defined herein. 20          The invention also provides processes, reagents and methods for increasing the efficiency of a biochemical reaction in vivo. Accodingly, any reference to a process for performing a biochemical reaction in an aqueous in vitro reaction system as used herein using any of the IDR reagents as defined herein may alternatively be defined as a process for performing a biochemical reaction in an aqueous in vivo reaction system 25 using any of the IDR reagents as defined herein. The invention provides any non-naturally occurring IDR-macromolecule or IDR-polypeptide described or defined herein for use in therapy, for use as a therapeutic, for use as a medicament, for use as a pharmaceutical agent or for use as a diagnostic agent. 2026234101   18 Sep 2026 The invention provides any non-naturally occurring IDR-macromolecule or IDR-polypeptide described or defined herein for use in a method for the treatment of the human or animal body by therapy. The invention provides any non-naturally occurring IDR-macromolecule or 5 IDR-polypeptide described or defined herein for use in a diagnostic method practised on the human or animal body. The invention provides any non-naturally occurring IDR-macromolecule or IDR-polypeptide described or defined herein for use in the manufacture of a medicament for the treatment of the human or animal body by therapy. 10          The invention provides any non-naturally occurring IDR-macromolecule or IDR-polypeptide described or defined herein for use in the manufacture of a diagnostic agent for a diagnostic method practised on the human or animal body. The invention provides a method of treatment of a human or animal comprising administering a therapeutically effective amount of any non-naturally occurring IDR- 15 macromolecule or IDR-polypeptide described or defined herein to a human or animal in need thereof. In any one of the above-described processes, reagents and methods for increasing the efficiency of a biochemical reaction, the non-naturally occurring IDR-macromolecule or IDR-polypeptide is capable of promoting liquid-liquid demixing. 20 Said liquid-liquid demixing may be capable of promoting the formation of phase-separated aqueous compartments in solution, including detectable phase-separated aqueous particles in solution. Said liquid-liquid demixing or said formation of detectable phase-separated compartments or particles thereby increases the efficiency of the biochemical reaction caused by the IDR-macromolecule or the IDR-polypeptide. 25          A process for performing an in vitro biochemical reaction as used herein includes any biochemical reaction performed in vitro in solution in a reaction vessel or within a host cell in culture by introducing an IDR reagent as defined herein into the solution or introducing or expressing an IDR reagent in a cultured host cell to promote liquid-liquid demixing in the solution or in the cultured host cell. In any such 30 biochemical reaction, the liquid-liquid demixing in the solution or in the cultured host 2026234101   18 Sep 2026 cell promotes phase separation in the solution or in the cultured host cell, as described and defined herein. Any such biochemical reaction may be performed in order to assess the efficacy of any IDR amino acid sequence as described and defined herein in promoting liquid-5 liquid demixing in the solution or in the cultured host cell and / or in promoting phase separation in the solution or in the cultured host cell. Any such biochemical reaction may be performed in order to assess the efficacy of a test agent, such as a drug, a polypeptide or any other molecule, in stimulating or enhancing liquid-liquid demixing mediated by the IDR amino acid sequence in the 10 solution or in the cultured host cell and / or in stimulating or enhancing phase separation mediated by the IDR amino acid sequence in the solution or in the cultured host cell, preferably wherein the test agent interacts with the IDR amino acid sequence. Any such biochemical reaction may be performed in order to assess the efficacy of a test agent, such as a drug, a polypeptide or any other molecule, in inhibiting liquid-15   liquid demixing mediated by the IDR amino acid sequence in the solution or in the cultured host cell and / or in inhibiting phase separation mediated by the IDR amino acid sequence in the solution or in the cultured host cell, preferably wherein the test agent interacts with the IDR amino acid sequence. Any of the processes described herein for performing an in vitro, in vivo or ex 20 vivo biochemical reaction may exclude a process for cloning a human being. Any of the processes described herein for performing an in vitro, in vivo or ex vivo biochemical reaction may exclude a process for modifying the germ line genetic identity of a human being. Any of the processes described herein for performing an in vitro, in vivo or ex 25 vivo biochemical reaction may exclude a process involving the use of a human embryo, or the use of a totipotent human cell. Any host cell described herein may exclude a human embryo, or a totipotent human cell, or a human germ line cell. Whilst embracing in vivo use in some aspects, the invention embraces the 30 exclusion of in vivo use in some aspects. Accordingly, any of the processes, uses or 2026234101   18 Sep 2026 methods etc. described herein for performing a biochemical reaction in an aqueous reaction system may exclude an in vivo aqueous reaction system. Whilst embracing ex vivo use in some aspects, the invention embraces the exclusion of ex vivo use in some aspects. Accordingly, any of the processes, uses or 5 methods etc. described herein for performing a biochemical reaction in an aqueous reaction system may exclude an ex vivo aqueous reaction system. In any of the methods, processes, uses or IDR reagents described herein, the efficiency of the reaction in the system may be increased by the IDR-macromolecule or the IDR-polypeptide compared to the efficiency of the reaction in the system following 10   the introduction of the at least one macromolecule or polypeptide under the same reaction conditions except that the at least one macromolecule or polypeptide does not comprise one or more functional intrinsically disordered regions (IDRs). In any of the methods, processes, uses or IDR reagents described herein involving at least one macromolecule or at least one polypeptide tagged with an amino 15   acid sequence comprising or consisting of one or more functional intrinsically disordered regions (IDRs) (IDR-tagged macromolecule or IDR-tagged polypeptide) the efficiency of the reaction in the system may be increased by the IDR-tagged macromolecule or IDR-taggged polypeptide compared to the efficiency of the reaction in the system following the introduction of the at least one macromolecule or 20 polypeptide under the same reaction conditions except that the at least one macromolecule or polypeptide has not been tagged with the amino acid sequence comprising or consisting of the one or more functional IDRs. Accordingly, whether an IDR-macromolecule or IDR-polypeptide, or an IDR-tagged macromolecule or IDR-tagged polypeptide, is capable of increasing the 25 efficiency of the reaction in the system can be established very simply by comparing the reaction efficiency of the macromolecule or polypeptide with or without the one or more functional IDRs. The skilled person is able to carry out simple comparative tests to establish the relevant functional capability. Exemplary test assays are described further herein. 30          Similarly, whether an IDR-macromolecule or IDR-polypeptide, or an IDR- tagged macromolecule or IDR-tagged polypeptide, is capable of causing molecules 2026234101   18 Sep 2026 necessary for the performance of the reaction to co-localise with the IDR-macromolecule or the IDR-polypeptide, or with the IDR-tagged macromolecule or IDR-tagged polypeptide, within the plurality of phase-separated aqueous compartments, or to further stimulate or enhance co-localisation of molecules necessary for the performance 5 of the reaction within the plurality of phase-separated aqueous compartments, thereby increasing the efficiency of the biochemical reaction in the system, can also be established very simply by comparing co-localisation with or without the one or more functional IDRs. Again, the skilled person is able to carry out simple comparative tests to establish the relevant functional capability. Exemplary test assays are described 10    further herein. Similarly whether providing multivalent metal ions to an IDR-macromolecule or an IDR-polypeptide, or to an IDR-tagged macromolecule or IDR-tagged polypeptide thereby further stimulating or enhancing liquid-liquid demixing and the formation of the plurality of phase-separated aqueous compartments and thereby further increasing the 15   efficiency of the biochemical reaction in the system can also be established very simply by comparing liquid-liquid demixing with or without providing multivalent metal ions. Again, the skilled person is able to carry out simple comparative tests to establish the relevant functional capability. Exemplary test assays are described further herein. Similarly whether providing ATP to an IDR-macromolecule or an IDR- 20 polypeptide, or to an IDR-tagged macromolecule or IDR-tagged polypeptide may further simulate or enhance liquid-liquid demixing and the formation of the plurality of phase-separated aqueous compartments and thereby further increase the efficiency of the biochemical reaction in the system can also be established very simply by comparing liquid-liquid demixing with or without providing ATP. Whether providing 25 ATP to an IDR-macromolecule or an IDR-polypeptide, or to an IDR-tagged macromolecule or IDR-tagged polypeptide to further stimulate or enhance colocalisation of molecules necessary for the performance of the reaction within the plurality of phase-separated aqueous compartments, thereby increasing the efficiency of the biochemical reaction in the system, can also be established very simply by 30 comparing co-localisation with or without providing ATP. Again, the skilled person is 2026234101   18 Sep 2026 able to carry out simple comparative tests to establish the relevant functional capability. Exemplary test assays are described further herein. Assays are described herein to establish the capability to cause liquid-liquid demixing by reference to the capability to cause the formation of phase-separated 5 aqueous particles (see e.g. “phase separation assay method” as described herein). The same assay can be used to establish the capability to cause co-localisation of molecules necessary for the performance of the reaction within the phase-separated aqueous compartments (particles). Assays are described herein to establish the capability to increase the efficiency of a reaction by reference to the capability to increase the 10   efficiency of an RPA method (see e.g. “RPA assay method” as described herein). Such an assay can be used to evaluate the capability of an amino acid sequence consisting of or comprising one or more functional intrincically disordered regions (IDRs) to increase the efficiency of a reaction, and / or to evaluate the capability of divalent metal ions to further increase the efficiency of a reaction and / or to evaluate the capability of ATP to 15    further increase the efficiency of a reaction. Using simple assays as described herein the skilled person is able to deterime an increase in the efficiency of a reaction of 5% or more, the increase in the efficiency of the reaction may be 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, 50% or more, 55% or more, 60% or 20 more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, 95% or more or 100% or more. Crowding agent 25          A crowding agent is typically a high molecular weight macromolecule, such as a protein or a synthetic block polymer. A crowding agent is considered to be essentially biochemically inert, i.e. it does not contribute to a specific interaction or catalysis. It is widely postulated that a crowding agent exerts an influence on a biological / biochemical system, be it an in vitro or an in vivo system, through the effects 30 of its physical occupation of volume in a solution thus causing steric obstruction and a reduction in available open solvent space. By this excluded volume mechanism 2026234101   18 Sep 2026 crowding agents appear to increase the effective concentration of other macromolecules, with particular influence on altering dissociation constants and favouring the association of interacting macromolecules such as multiple proteins which come together into specific organised complexes. The size of the crowding effect depends in particular on 5 the molecular mass of the molecules involved, being generally much stronger with larger molecules. Thus, as a general rule, macromolecular crowding is an effect exerted by large molecules on the properties of other large molecules. Furthermore, crowding agents have been widely described as being capable of facilitating the formation of biological / biochemical systems with preferred phases in 10   which reactants segregate themselves into micron-sized phase-separated particles. This effect arises substantially from the effects of volume exclusion on the dissociation constant of macromolecular complexes which become relatively “confined” due to the inability to readily diffuse into the largely volume-occupied bulk solvent. Additionally and / or alternatively, some crowding agents such as block chain polymers like 15   polyethylene glycol may exhibit kosmotropic character which leads them to exert an overall alteration in the structure of bulk water, typically lowering water density. Such changes in bulk solvent characteristics may also exert complex effects on other macromolecules and their assemblies whose surfaces must interact with water. This in turn may also promote segregation of those other macromolecules into an alternate 20 phase, significantly enriched in the biological components and concomitantly depleted in the crowding agent which principally occupies the bulk solvent phase. In either scenario, either by simple volume occupation or solvent modification, the effects individually or in combination of crowding agents in stimulating the condensation of macromolecules into phase-distinct condensates appears to operate by a 25 “repulsive” rather than by an “attractive” mechanism from the perspective of the condensate components. In other words, from the perspective of the components that are highly enriched in the condensate the crowding agent acts to create a bulk phase environment that cannot be readily penetrated by diffusion, and / or whose bulk solvent character is modified in a manner that it presents a net enthalpic disadvantage to 30 disperse into. It is in this manner that the effects of high concentrations of crowding agents, typically greater than 1% w / v, are referred to herein in stimulating phase 2026234101   18 Sep 2026 separation by functioning through an “obstructive” or “repulsive” mechanism insofar as the phenomenon arises due to the condensate components being unable to readily disperse as they would in the absence of the crowding agent. However, at the same time, in view of its generally inert properties, the crowding agent has little or no direct 5 debilitating effects on other specific molecules in the system, for example because the crowding agent does not significantly interact with or exert effects on specific molecular side-chains in a direct fashion. In standard RPA reactions polyethylene glycols (PEGs) can have profound effects on recombination / DNA synthesis. PEGs can influence the number of multiple 10 invasion / extension cycles that occur, for example, when RecA is combined with Gp32. PEGs can stimulate amplification reactions configured in several different ways. PEGs and other similar crowding agents may affect the cooperativity of Gp32 and recombinases, they may affect polymerase processivity and they may affect the hybridisation rate and behaviour of oligonucleotides in solution. The chain length of 15 the polyethylene glycol can influence results. PEGs may also increase the stability of recombinase-loaded filaments and the increased persistence may increase RPA efficacy. To exert its effects in an in vitro biochemical reaction milieu an added crowding agent is typically present at concentrations at which steric exclusion / confmement effects are predicted to occur, typically above approximately 1% by volume of the reaction, or 20 by weight of the reaction. In standard RPA reactions a crowding agent is present at concentrations of approximately 1% to 12% by volume of the reaction or by weight of the reaction. The terms “macromolecular crowding agent” or more simply “crowding agent” are very well recognised and art-understood terms. This is apparent from the literature 25 in which the terms are used extensively. For example Kuznetsova, I., M. et al., (What Macromolecular Crowding Can Do to a Protein, 2014, Int. J. Mol. Sci., 15, pp 2309023140) provide a review which purports to cover more than 320 papers and is suggested to represent one of the most comprehensive compendia of the current knowledge in the field. The term “crowding agent” is used extensively throughout the text highlighting 30 its ubiquitous use (see also Mixed Macromolecular Crowding: A Protein and Solvent Perspective, Biswas, S. et al., 2018, ACS Omega, 3(4), pp4316-4330 and Common 2026234101   18 Sep 2026 Crowding Agents Have Only a Small Effect on Protein-Protein Interactions, Phillip Y. et al., 2009, Biophysical Journal, 97 pp875-885 875). A compound or macromolecule can be identified as a crowding agent by means known in the art. For example a crowding agent can be identified as such via its 5 experimentally determined and calculated hydrodynamic radius (Kuznetsova et al., supra). A crowding agent can be identified as such via sol-gel glass encapsulation analysis (Kuznetsova et al., supra). The following compounds are examples of known crowding agents. A synthetic block polymer , a polyethylene glycol (PEG), PEG 1450, PEG 2050, PEG3000, PEG 10   4600, PEG 6000, PEG 8000, PEG 10000, PEG 20000, PEG 35000, PEG compound molecular weight 15,000 to 20,000 (also known as Carbowax 20M), a dextran, Dextran 6, Dextran 40, Dextran 70, Dextran 670, Dextran sulfate 10, Dextran sulfate 500, a ficoll, Ficoll 70, Ficoll 400, Poly(sodium 4-styrene sulfonate) (PSS), Bovine pancreatic trypsin inhibitor (BPTI), Ribonuclease A, Lysozyme, |3-Lac to globulin, Hemoglobin, 15 Bovine serum albumin (BSA). In any one of the methods, processes and uses of the present invention, including in any one of the RPA methods, processes and uses of the present invention, the methods, processes and uses may be performed in the absence of a crowding agent. In any one of the methods, processes and uses of the present invention, including 20 in any one of the RPA methods, processes and uses of the present invention, the methods, processes and uses may be performed in the presence of a crowding agent. In any one of the methods, processes and uses of the present invention, including in any one of the RPA methods, processes and uses of the present invention, the methods, processes and uses may be performed in the presence of a crowding agent, 25 wherein the crowding agent is provided at a concentration which provides an enhancement of the increase in the efficiency of the biochemical reaction which is provided by the IDR-macromolecule or the IDR-polypeptide. In any one of the methods, processes and uses of the present invention, including in RPA methods, processes and uses of the present invention, the methods, processes 30 and uses may be performed in the presence of a crowding agent, wherein the crowding agent is provided at a concentration which provides an additive effect on the efficiency 2026234101   18 Sep 2026 of the biochemical reaction which is provided by the IDR-macromolecule or the IDR-polypeptide. In any one of the methods, processes and uses of the present invention, including in any one of the RPA methods, processes and uses of the present invention, the 5 methods, processes and uses may be performed in the presence of a crowding agent, wherein the crowding agent is provided at a concentration which provides a synergistic effect on the efficiency of the biochemical reaction which is provided by the IDR-macromolecule or the IDR-polypeptide. In any one of the methods, processes and uses of the present invention, including 10 in any one of the RPA methods, processes and uses of the present invention, the methods, processes and uses may be performed in the presence of a crowding agent, wherein the introduction of the IDR-macromolecule or the IDR-polypeptide into the biochemical reaction system lowers the concentration of crowding agent which would be required to achieve the same increase in the efficiency of the biochemical reaction in 15   the absence of the introduction of the IDR-macromolecule or the IDR-polypeptide into the biochemical reaction system. In any one of the above-described methods, processes and uses which may be performed in the presence of a crowding agent, the crowding agent may be present at a concentration below that at which its normal biological effects (steric 20 exclusion / confinement effects) occur. In any one of the above-described methods, processes and uses which may be performed in the presence of a crowding agent, the crowding agent may be present at concentrations below approximately 3% by volume of the reaction or by weight of the reaction, approximately 2% by volume of the reaction or by weight of the reaction, 25 approximately 1% by volume of the reaction or by weight of the reaction or approximately 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2% or 0.1% by volume of the reaction or by weight of the reaction. If used in any one of the methods, processes and uses of the invention, including RPA reaction methods, any suitable crowding agent may be used. Examples of suitable 30 crowding agents are provided herein. 2026234101   18 Sep 2026 Macromolecule or polypeptide comprising an intrinsically disordered region (IDR) The methods, processes and reagents of the present invention involve “IDR-macromolecules”, including “IDR-tagged macromolecules”, as described herein. The 5 methods, processes and reagents of the present invention involve “IDR-polypeptides”, including “IDR-tagged polypeptides”, as described herein. Any such IDR-macromolecules, IDR-tagged macromolecules, IDR-polypeptides or IDR-tagged polypeptides may be referred to interchangeably herein as IDR reagents or IDR-based reagents. 10         An IDR-macromolecule or IDR-polypeptide or an IDR-tagged macromolecule or IDR- tagged polypeptide as used herein is any macromolecule, or polypeptide or protein, which comprises one or more functional intrinsically disordered regions (IDRs). An IDR-macromolecule or IDR-polypeptide or an IDR-tagged macromolecule 15 or IDR-tagged polypeptide as used herein is any macromolecule, or polypeptide or protein, which comprises an amino acid sequence consisting of or comprising one or more functional intrinsically disordered regions (IDRs). Accordingly, an IDR-macromolecule or IDR-polypeptide, as referred to herein, may thus refer to: a macromolecule or polypeptide which comprises an amino acid 20 sequence consisting of one or more functional intrinsically disordered regions; or a macromolecule or polypeptide which comprises an amino acid sequence comprising one or more functional intrinsically disordered regions. In addition, an IDR-macromolecule comprising one or more functional intrinsically disordered regions (IDRs), as referred to herein may be a macromolecule of 25 interest tagged with an amino acid sequence consisting of or comprising one or more functional intrinsically disordered regions (IDRs). Such an IDR-tagged macromolecule is also an IDR reagent as defined herein. An IDR-tagged polypeptide comprising one or more functional intrinsically disordered regions (IDRs), as referred to herein, may be a polypeptide of interest tagged with an amino acid sequence consisting of or comprising 30 one or more functional intrinsically disordered regions (IDRs). Such an IDR-tagged polypeptide is also an IDR reagent as defined herein. 2026234101   18 Sep 2026 An IDR-tagged macromolecule or IDR-tagged polypeptide, as used herein, is any macromolecule, or polypeptide or protein, which is “tagged” with an amino acid sequence consisting of or comprising one or more functional intrinsically disordered regions (IDRs). 5          Accordingly, an IDR-tagged macromolecule or IDR-tagged polypeptide, as referred to herein, may thus refer to: a macromolecule or polypeptide which is tagged with an amino acid sequence consisting of one or more functional intrinsically disordered regions; or a macromolecule or polypeptide which is tagged with an amino acid sequence comprising one or more functional intrinsically disordered regions. 10          A tagged amino acid sequence consisting of or comprising one or more functional intrinsically disordered regions (IDRs) is not found naturally or ordinarily in the macromolecule or polypeptide or protein to which it is tagged at the tagged position. Accordingly, a tagged amino acid sequence consisting of or comprising one or more functional intrinsically disordered regions (IDRs) can be considered to be an exogenous 15   amino acid sequence compared to the macromolecule or polypeptide or protein to which it is tagged. The tagged macromolecule or polypeptide or protein can therefore be considered to be a non-naturally occurring, artificial or genetically engineered macromolecule, polypeptide or protein. Mechanisms by which amino acid sequences may be “tagged” to polypeptides 20 and other macromolecules are explained further herein. Any one or more functional intrinsically disordered regions (IDRs) may be tagged to a macromolecule or polypeptide or protein, including any one or more of the specific IDR amino acid tag sequences disclosed herein, or any one or more functional variants, analogs, homologs or derivatives thereof. 25          For use in the present invention both an intrinsically disordered region polypeptide sequence and a domain thereof should be “functional”. The term “functional” means that any IDR amino acid sequence must possess one of the functional properties outlined further herein. The term “intrinsically disordered region” is an art-understood term used 30 commonly in the technical field. For a comprehensive review see: Classification of 2026234101   18 Sep 2026 Intrinsically Disordered Regions and Proteins, van der Lee et al., 2014, Chern. Rev. 114,pp 6589-6631. The invention provides, inter alia, a process of performing a biochemical reaction in an aqueous in vitro reaction system, wherein the biochemical reaction is 5 dependent on the function of at least one reaction macromolecule, optionally at least one reaction polypeptide, the process comprising: introducing at least one IDR-macromolecule into the in vitro reaction system under conditions suitable for performing the reaction, wherein the at least one IDR-macromolecule comprises one or more functional intrinsically disordered regions (IDRs), wherein upon introduction of 10   the at least one IDR-macromolecule into the in vitro reaction system the efficiency of the biochemical reaction is increased by the at least one IDR-macromolecule. The efficiency of the biochemical reaction is increased by the one or more functional IDRs of the IDR-macromolecule. In any such process the at least one IDR-macromolecule may be at least one IDR-polypeptide. In any such process, the IDR-macromolecule or 15 IDR-polypeptide comprising one or more functional intrinsically disordered regions (IDRs) may not be the “reaction macromolecule” or “reaction polypeptide” upon whose function the biochemical reaction depends. Thus in any such process the IDR-macromolecule or IDR-polypeptide may have no intrinsic biochemical role in the biochemical reaction per se. Nevertheless, its introduction into the reaction system 20 leads to an increase in the efficiency of the biochemical reaction. The process of performing a biochemical reaction in an in vitro reaction system may be a process wherein the biochemical reaction is dependent on the function of the at least one IDR-macromolecule, wherein upon its introduction into the in vitro reaction system the at least one IDR-macromolecule performs its reaction function in the 25 biochemical reaction and increases the efficiency of the reaction. In any such process the at least one IDR-macromolecule maybe at least one IDR-polypeptide. In any such process, the IDR-macromolecule or IDR-polypeptide does have an intrinsic biochemical role in the biochemical reaction per se. Thus the at least one IDR-macromolecule or IDR-polypeptide comprising one or more functional intrinsically disordered regions 30 (IDRs) is the “reaction macromolecule” or “reaction polypeptide” upon whose function the biochemical reaction depends. 2026234101   18 Sep 2026 The at least one IDR-macromolecule or IDR-polypeptide comprises an amino acid sequence comprising or consisting of one or more functional intrinsically disordered regions. The IDR-macromolecule or IDR-polypeptide is introduced into the biochemical reaction system under conditions suitable for performing the biochemical 5 reaction. Because of the presence of the one or more functional intrinsically disordered regions, the IDR-macromolecule or IDR-polypeptide increases the efficiency of the reaction. By increasing the efficiency of the reaction it is meant that the efficiency of the reaction is improved compared to the efficiency of the reaction which would be 10   observed if the IDR-macromolecule or IDR-polypeptide was provided without an amino acid sequence comprising or consisting of the one or more functional intrinsically disordered regions. Such an improvement can readily be established by comparative testing of the reaction macromolecule or polypeptide with and without the IDR amino acid sequence. 15          In any one of the methods of the invention the one or more functional intrinsically disordered regions of the IDR-macromolecule or IDR-polypeptide promotes / causes liquid-liquid demixing in the reaction system leading to phase separation. The functional capability of promoting liquid-liquid demixing leading to phase separation in a reaction system can readily be established, for example by 20 performing a phase separation assay as described herein. Such liquid-liquid demixing promotes phase separation and this may lead to the formation of phase separated compartments in the reaction system, such as particles which are detectable, e.g. under microscopic observation, as detailed further herein. An IDR-macromolecule or IDR-polypeptide may or may not posess catalytic 25 activity. For example, an IDR-polypeptide may have catalytic activity, such as a polymerase enzyme used in a recombinase polymerase amplification reaction, as described further herein. An IDR-polypeptide may not have catalytic activity, such as a single strand stabilizing agent used in a recombinase polymerase amplification reaction, e.g. Gp32 as described further herein. 30          As discussed further herein, whether an IDR-macromolecule or IDR-polypeptide has catalytic activity or not, the IDR-macromolecule or IDR-polypeptide may possess a 2026234101   18 Sep 2026 function which is required for or influences the biochemical reaction, such that in the absence of the IDR-macromolecule or IDR-polypeptide in the biochemical reaction system the biochemical reaction cannot proceed or proceeds with reduced efficiency. Alternatively, as discussed further herein, the IDR-macromolecule or IDR-polypeptide 5 may not possess any function which is required for or which influences the biochemical reaction per se. Nevertheless, because of the IDR amino acid sequences, the introduction of the IDR-macromolecule or IDR-polypeptide into the biochemical reaction system leads to an increase in the efficiency of the biochemical reaction compared to the efficiency observed in the absence of the IDR-macromolecule or IDR-10 polypeptide, or in the presence of the same macromolecule or polypeptide without the IDR amino acid sequences. Structural properties of an IDR polypeptide 15          The presence of IDRs in amino acid sequences may readily be determined by structural analysis. A large number of bioinformatics-based platforms are available for the prediction of the presence of IDRs within polypeptides and proteins. These include ELM, MiniMotif, SLiMPrints, phylo-HMM, DiliMot, SLiMFinder, Phospho.ELM, PhosphoSite, PHOSIDA, ScanSite, NetPhorest, NetworKIN, PhosphoNET, IDEAL, 20 MoRFpred, ANCHOR, Pfam, FFPred, DisProt, D2P2, and MetaDisorder. Any of these methods may be used to identify IDR amino acid sequences. If necessary, such IDR amino acid sequences can be tested to assess their functional properties, as described futher herein. A preferred bioinformatics-based platform for IDR amino acid sequence 25 identification is the MetaDisorder software program (MetaDisorder: a meta-server for the prediction of intrinsic disorder in proteins. Kozlowski, L. P., et al., BMC Bioinformatics, 2012, 13(1): 111). The MetaDisorder program is available freely online (http: / / genesilico.pl / metadisorder / ). Using this program an amino acid sequence of 30 interest is simply pasted into an internet browser window and the program is initiated. 2026234101   18 Sep 2026 As the online documentation explains, any amino acid region which scores >0.5 in the software package is considered to comprise an intrinsically disordered region. Using the MetaDisorder software platform the inventors have identified a number of amino acid sequences which comprise one or more intrinsically disordered 5 regions. These are set out in Table 1. Accordingly, in any one of the methods, processes and uses, or in any one of the non-naturally occurring IDR-macromolecules, IDR-fusion macromolecules or isolated nucleic acid molecules encoding the same, recombinant polynucleotide expression vectors or host cells, the one or more functional IDRs of the IDR-macromolecule or the 10 IDR-polypeptide may be characterised as a sequence of amino acids which scores greater than 0.5 when analysed by the algorithm MetaDisorder. The sequence of amino acids may be a sequence of amino acids which scores greater than 0.5 when analysed by the algorithm MetaDisorder accordiong to the methods of Kozlowski, L. P., et al., BMC Bioinformatics, 2012, 13(1): 111. 15          The invention provides and relates to preferred IDR amino acid sequences which comprise or consist of the amino acid sequence of any one of SEQ ID NOs 1 to 43 (Table 1) and variants thereof. In all cases a variant of an amino acid sequence of any one of SEQ ID NOs 1 to 43 is a functional variant which retains the IDR functional properties, as described further herein. 20          Furthermore, as described further herein, the IDR-macromolecule or IDR- polypeptide may comprise or consist of a macromolecule or polypeptide tagged with an amino acid sequence which comprises or consists of the amino acid sequence of any one of SEQ ID NOs 1 to 43, or which comprises or consists of a functional variant amino acid sequence of SEQ ID NOs 1 to 43. 25          The functional variant may have at least 80% sequence identity compared to the herein-described IDR amino acid sequence (Table 1). The functional variant may have at least 81 % sequence identity compared to the herein-described IDR amino acid sequence (Table 1), or 82% sequence identity, or 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity. 30           For the purpose of this invention, in order to determine the percent identity e.g. between an amino acid sequence of any one of SEQ ID NOs 1 to 43 and a functional 2026234101   18 Sep 2026 variant of an amino acid sequence of any one of SEQ ID NOs 1 to 43 the two respective amino acid sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in a first sequence for optimal alignment with a second sequence). The nucleotide residues at nucleotide positions are then compared. When a position in the 5 first sequence is occupied by the same nucleotide residue as the corresponding position in the second sequence, then the nucleotides are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences (i.e., % identity = number of identical positions / total number of positions in the reference sequence x 100). 10          Typically the sequence comparison is carried out over the full length of the reference sequence. For example, if the skilled person wished to determine whether a given (“variant”) sequence is 80% identical to SEQ ID NO. 2, SEQ ID NO. 2 would be the reference sequence. For example, to assess whether a variant sequence is at least 80% identical to SEQ ID NO. 2 (an example of a reference sequence), the skilled person 15 would carry out an alignment over the length of SEQ ID NO. 2, and identify how many positions in the test sequence were identical to those of SEQ ID NO. 2. If at least 80% of the positions are identical, the test sequence is at least 80% identical to SEQ ID NO. 2. If the sequence is shorter than SEQ ID NO. 2, the gaps or missing positions should be considered to be non-identical positions. 20          The skilled person is aware of different computer programs that are available to determine the homology or identity between two sequences. For instance, a comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. The percent identity between two amino acid or nucleic acid sequences may be determined, for example, using the Needleman 25 and Wunsch (1970) algorithm which has been incorporated into the GAP program in the Accelrys GCG software package (available at http: / / www.accelrys.com / products / gcg / ), using either a Blosum 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6. A functional variant of an amino acid sequence of any one of SEQ ID NOs 1 to 30   43 may be an amino acid sequence that differs by having a number of amino acids less compared to the amino acid sequence of any one of SEQ ID NOs 1 to 43 respectively 2026234101   18 Sep 2026 (i.e. the functional variant is shorter), or that differs by having a number of amino acids more compared to the amino acid sequence of any one of SEQ ID NOs 1 to 43 respectively (i.e. the functional variant is longer). The functional variant may thus contain one or more amino acid deletions and / or one or more insertions compared to the 5 reference amino acid sequence. The number of amino acids in the functional variant amino acid sequence by which the variant differs from the reference sequence can be 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, 13 or more, 14 or more, 15 or more, 16 or more, 17 or more, 18 or more, 19 or more or 20 or more. 10          A functional variant of an amino acid sequence of any one of SEQ ID NOs 1 to 43 may for example comprise conservative amino acid substitutions of amino acid residues indicated in the sequences listed in Table 1. Conservative substitutions may be made, for example according to the table below which describes a generally accepted grouping of amino acids. The functional variant may thus contain conservative amino 15 acid substitutions compared to the reference amino acid sequence. The number of amino acids in the functional variant amino acid sequence which are conservative amino acid substitutions compared to the reference sequence can be 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, 13 or more, 14 or more, 15 or more, 16 or more, 17 or 20 more, 18 or more, 19 or more or 20 or more. 2026234101   18 Sep 2026 Set Sub-set Hydrophobic FWYHKMILVAGC Aromatic F W YH Aliphatic ILV Polar WYHKREDCSTNQ Charged HKRED Positively charged HKR Negatively charged ED Small VCAGSPTND Tiny AGS Whether a given variant retains the IDR functional properties may readily be established, for example by methods described further herein. Table 1 - amino acid sequences comprising intrinsically disordered regions Tag Original protein Accession ID Organism Position Length Comment Sequence SEQ ID NO fib (or fib-1) rRNA 2'-o-methyltransfe rase fibrillarin Uniprot: P22087 Homo sapiens 3-45 43 N / A ProGlyPheSerProArgGlyGlyGlyPheGlyGlyArgGlyGlyP HeGlyAspArgGlyGlyArgGlyGlyArgGlyGlyPheGlyGlyGl yArgGlyArgGlyGlyGlyPheArgGlyArgGlyArg 1 fib2 rRNA 2'-0-methyltransfe rase fibrillarin Uniprot: P22087 Homo sapiens 3-45 86 double of fib tag ProGlyPheSerProArgGlyGlyGlyPheGlyGlyArgGlyGlyP heGlyAspArgGlyGlyArgGlyGlyArgGlyGlyPheGlyGlyGl yArgGlyArgGlyGlyGlyPheArgGlyArgGlyArgProGlyPhe SerProArgGlyGlyGlyPheGlyGlyArgGlyGlyPheGlyAspA rgGlyGlyArgGlyGlyArgGlyGlyPheGlyGlyGlyArgGlyAr gGlyGlyGlyPheArgGlyArgGlyArg 2 Fib [short] rRNA 2'-0-methyltransfe rase fibrillarin Uniprot: P22087 Homo sapiens 8-32 25 N / A ArgGlyGlyGlyPheGlyGlyArgGlyGlyPheGlyAspArgGlyG lyArgGlyGlyArgGlyGlyPheGlyGly 3 fib-2 (clipperl) N / A N / A N / A N / A 36 selfdesigned: 7 amino acids deletion at the C— terminus of fib (fib-1) ProGlyPheSerProArgGlyGlyGlyPheGlyGlyArgGlyGlyP HeGlyAspArgGlyGlyArgGlyGlyArgGlyGlyPheGlyGlyGl yArgGlyArgGlyGly 4 fib-3 (clipper2) N / A N / A N / A N / A 27 selfdesigned: 16 amino acids ProGlyPheSerProArgGlyGlyGlyPheGlyGlyArgGlyGlyP HeGlyAspArgGlyGlyArgGlyGlyArgGlyGly 5 WO 2021 / 094746                                                 PCT / GB2020 / 052866 deletion at the C-terminus of fib (fib-1) fib-4 (clipper3) N / A N / A N / A N / A 15 selfdesigned: 28 amino acids deletion at the C-terminus of fib (fib-1) ProGlyPheSerProArgGlyGlyGlyPheGlyGlyArgGlyGly 6 PCF Protein PCF11 Uniprot: P39081 Saccharomyc es cerevisiae 208-287 67 modificat ion : amino acids deletion from originial positions 234-246 GlnValGlnMetGlnLeuArgGlnValPheSerGlnAspGlnGlnV alLeuGlnGluArgMetArgTyrHisGluLeuGlnGlnGlnGlnGl nGlnGlnTyrHisGluThrLysAspMetValGlySerTyrThrGln AsnSerAsnSerAlalleProLeuPheGlyAsnAsnSerAspThrT hrAsnGlnGlnAsnSer 7 ENT1 Ent2p Uniprot: C7GIT6 Saccharomyc es cerevisiae 530-588 59 N / A GlnAsnProThrGlylleSerTyrSerGlnProGlnGlnGlnGlnG InProGlnGlnGlnProGlnTyrMetGlnAsnPheGlnGlnGlnGl nProGlnTyrAlaGlnAsnPheGlnGlnGlnProGlnTyrThrGln AsnTyrGlnGlnGlnProGlnTyrlleGlnProHisGln 8 HRP1 Nuclear polyadenylate d RNA-binding protein 4 Uniprot: Q99383 Saccharomyc es cerevisiae 331-388 58 N / A GlyGlyAsnAsnGlyGlyAsnAsnMetAsnArgArgGlyGlyAsnP heGlyAsnGlnGlyAspPheAsnGlnMetTyrGlnAsnProMetMe tGlyGlyTyrAsnProMetMetAsnProGlnAlaMetThrAspTyr TyrGlnLysMetGlnGluTyrTyrGlnGlnMetGln 9 WO 2021 / 094746                                                 PCT / GB2020 / 052866 HRP2 Nuclear polyadenylate d RNA-binding protein 4 Uniprot: Q99383 Saccharomyc es cerevisiae 331-388 116 double of HRP1 tag GlyGlyAsnAsnGlyGlyAsnAsnMetAsnArgArgGlyGlyAsnP heGlyAsnGlnGlyAspPheAsnGlnMetTyrGlnAsnProMetMe tGlyGlyTyrAsnProMetMetAsnProGlnAlaMetThrAspTyr TyrGlnLysMetGlnGluTyrTyrGlnGlnMetGlnGlyGlyAsnA snGlyGlyAsnAsnMetAsnArgArgGlyGlyAsnPheGlyAsnGl nGlyAspPheAsnGlnMetTyrGlnAsnProMetMetGlyGlyTyr AsnProMetMetAsnProGlnAlaMetThrAspTyrTyrGlnLysM etGlnGluTyrTyrGlnGlnMetGln 10 DDX Probable atp-dependent RNA helicase DDX4 Uniprot: Q9NQI0 Homo sapiens 4-240 240 N / A MetGlyAspGluAspTrpGluAlaGlulleAsnProHisMetSerS erTyrValProIlePheGluLysAspArgTyrSerGlyGluAsnGl yAspAsnPheAsnArgThrProAlaSerSerSerGluMetAspAsp GlyProSerArgArgAspHisPheMetLysSerGlyPheAlaSerG lyArgAsnPheGlyAsnArgAspAlaGlyGluCysAsnLysArgAs pAsnThrSerThrMetGlyGlyPheGlyValGlyLysSerPheGly AsnArgGlyPheSerAsnSerArgPheGluAspGlyAspSerSerG lyPheTrpArgGluSerSerAsnAspCysGluAspAsnProThrAr gAsnArgGlyPheSerLysArgGlyGlyTyrArgAspGlyAsnAsn SerGluAlaSerGlyProTyrArgArgGlyGlyArgGlySerPheA rgGlyCysArgGlyGlyPheGlyLeuGlySerProAsnAsnAspLe uAspProAspGluCysMetGlnArgThrGlyGlyLeuPheGlySer ArgArgProValLeuSerGlyThrGlyAsnGlyAspThrSerGlnS erArgSerGlySerGlySerGluArgGlyGlyTyrLysGlyLeuAs nGluGluVallleThrGlySerGlyLysAsnSerTrpLysSerGlu AlaGluGlyGlyGluSerSerAspThrGln 11 HNRNPA1 Heterogeneous nuclear ribonucleopro tein Al Uniprot: QOVACO Homo sapiens 187-320 134 N / A AlaSerAlaSerSerSerGlnArgGlyArgSerGlySerGlyAsnP HeGlyGlyGlyArgGlyGlyGlyPheGlyGlyAsnAspAsnPheGl yArgGlyGlyAsnPheSerGlyArgGlyGlyPheGlyGlySerArg GlyGlyGlyGlyTyrGlyGlySerGlyAspGlyTyrAsnGlyPheG lyAsnAspGlySerAsnPheGlyGlyGlyGlySerTyrAsnAspPh eGlyAsnTyrAsnAsnGlnSerSerAsnPheGlyProMetLysGly GlyAsnPheGlyGlyArgSerSerGlyProTyrGlyGlyGlyGlyG InTyrPheAlaLysProGlnAsnGlnGlyGlyTyrGlyValSerSe rSerSerSerSerTyrGlySerGlyArgArgPhe 12 WO 2021 / 094746                                                 PCT / GB2020 / 052866 PolIICTD DNA-directed RNA polymerase II subunit RPB1 Uniprot: P04050 Saccharomyc es cerevisiae 1521-1724 204 N / A SerProPheGlyAlaTyrGlyGluAlaProThrSerProGlyPheG lyValSerSerProGlyPheSerProThrSerProThrTyrSerPr oThrSerProAlaTyrSerProThrSerProSerTyrSerProThr SerProSerTyrSerProThrSerProSerTyrSerProThrSerP roSerTyrSerProThrSerProSerTyrSerProThrSerProSe rTyrSerProThrSerProSerTyrSerProThrSerProSerTyr SerProThrSerProSerTyrSerProThrSerProSerTyrSerP roThrSerProSerTyrSerProThrSerProSerTyrSerProTh rSerProSerTyrSerProThrSerProSerTyrSerProThrSer ProAlaTyrSerProThrSerProSerTyrSerProThrSerProS erTyrSerProThrSerProSerTyrSerProThrSerProSerTy rSerProThrSerProAsnTyrSerProThrSerProSerTyrSer ProThrSerProGlyTyrSerProGlySerProAlaTyrSerProL ysGlnAspGluGln 13 shortPolCTD DNA-directed RNA polymerase II subunit RPB1 Uniprot: P04050 Saccharomyc es cerevisiae 1636-1677 42 modificat ions : S1653GISI 667TIS167 4T ThrSerProSerTyrSerProThrSerProSerTyrSerProThrS erProGlyTyrSerProThrSerProAlaTyrSerProThrSerPr oThrTyrSerProThrSerProThrTyrSerPro 14 mimicl N / A N / A N / A N / A 49 selfdesigned: seven repeats of YDPTSPS (SEQ ID NO: 129) motifs mimiking the c-terminus of RNA polymeras e II TyrAspProThrSerProSerTyrAspProThrSerProSerTyrA spProThrSerProSerTyrAspProThrSerProSerTyrAspPr oThrSerProSerTyrAspProThrSerProSerTyrAspProThr SerProSer 15 WO 2021 / 094746                                                 PCT / GB2020 / 052866 miinic2 N / A N / A N / A N / A 49 selfdesigned: seven repeats of YSPTDPS (SEQ ID NO: 130) motifs mimiking the c-terminus Of RNA polymeras e II TyrSerProThrAspProSerTyrSerProThrAspProSerTyrS erProThrAspProSerTyrSerProThrAspProSerTyrSerPr oThrAspProSerTyrSerProThrAspProSerTyrSerProThr AspProSer 16 C-PolCTD DNA-directed RNA polymerase II subunit RPB1 Uniprot: P04050 Saccharomyc es cerevisiae 1642-1662 21 N / A ProThrSerProSerTyrSerProThrSerProSerTyrSerProT yrSerProAlaTyrSer 17 Sup Eukaryotic peptide chain release factor GTP-binding subunit Uniprot: P05453 Saccharomyc es cerevisiae 1-114 115 insertion of QY between 61 and 62; mutation : D66QIA67G deletion of Q70 MetSerAspSerAsnGlnGlyAsnAsnGlnGlnAsnTyrGlnGlnT yrSerGlnAsnGlyAsnGlnGlnGlnGlyAsnAsnArgTyrGlnGl yTyrGlnAlaTyrAsnAlaGlnAlaGlnProAlaGlyGlyTyrTyr GlnAsnTyrGlnGlyTyrSerGlyTyrGlnGlnGlyGlyTyrGlnG InTyrGlnTyrAsnProGlnGlyGlyTyrGlnGlnTyrAsnProGl nGlyGlyTyrGlnGlnTyrAsnProGlnGlyGlyTyrGlnGlnGln PheAsnProGlnGlyGlyArgGlyAsnTyrLysAsnPheAsnTyrA snAsnAsnLeuGlnGlyTyrGln 18 Supl N / A N / A N / A N / A o selfdesigned: single repeat of YNPQGGYQQ (SEQ ID NO: 19) TyrAsnProGlnGlyGlyTyrGlnGln 19 WO 2021 / 094746                                                 PCT / GB2020 / 052866 Ui Sup2 N / A N / A N / A N / A 18 selfdesigned: two repeat of YNPQGGYQQ (SEQ ID NO: 19) TyrAsnProGlnGlyGlyTyrGlnGlnTyrAsnProGlnGlyGlyT yrGlnGln 20 Sup3 N / A N / A N / A N / A 27 selfdesigned: three repeat of YNPQGGYQQ (SEQ ID NO: 19) TyrAsnProGlnGlyGlyTyrGlnGlnTyrAsnProGlnGlyGlyT yrGlnGlnTyrAsnProGlnGlyGlyTyrGlnGln 21 Sup4 N / A N / A N / A N / A 36 selfdesigned: four repeat of YNPQGGYQQ (SEQ ID NO: 19) TyrAsnProGlnGlyGlyTyrGlnGlnTyrAsnProGlnGlyGlyT yrGlnGlnTyrAsnProGlnGlyGlyTyrGlnGlnTyrAsnProGl nGlyGlyTyrGlnGln 22 Hisl Dual specificity tyrosine-phosphorylati on-regulated kinase 1A isoform 1 Uniprot: Q13627 Homo sapiens 593-623 31 N / A ProGlnGlnAsnAlaLeuHisHisHisHisGlyAsnSerSerHisH isHisHisHisHisHisHisHisHisHisHisHisGlyGlnGlnAl a 23 His2 Homeobox protein OTX1 Uniprot: P32242 Homo sapiens 273-308 36 N / A AlaGlyHisHisHisHisHisProHisAlaHisHisProLeuSerG InSerSerGlyHisHisHisHisHisHisHisHisHisHisGlnGl yTyrGlyGlySerGly 24 His3 Serine / threon ine-protein kinase NLK Uniprot: Q9UBE8 Homo sapiens 25-67 43 N / A AlaGlyHisHisHisHisHisHisHisHisLeuProHisLeuProP roProHisLeuHisHisHisHisHisProGlnHisHisLeuHisPr oGlySerAlaAlaAlaValHisProValGlnGlnHis 25 WO 2021 / 094746                                                 PCT / GB2020 / 052866 His4 Forkhead box protein G1 Uniprot: P55316 Homo sapiens 32-73 42 N / A AsnHisHisAlaSerHisGlyHisHisAsnSerHisHisProGlnH isHisHisHisHisHisHisHisHisHisHisProProProProAl aProGlnProProProProProGlnGlnGlnGln 26 His5 Transcription factor MafA Uniprot: Q8NHW3 Homo sapiens 145-220 76 N / A SerGlyHisHisGlyAlaHisHisGlyAlaHisHisProAlaAlaA laAlaAlaTyrGluAlaPheArgGlyProGlyPheAlaGlyGlyGl yGlyAlaAspAspMetGlyAlaGlyHisHisHisGlyAlaHisHis AlaAlaHisHisHisHisAlaAlaHisHisHisHisHisHisHisH IsHisHisGlyGlyAlaGlyHisGlyGlyGlyAlaGlyHisHis 27 His6 Transcription al repressor protein YY1 Uniprot: P25490 Homo sapiens 54-30 27 N / A GlyGlyGlyGlyAspHisGlyGlyGlyGlyGlyHisGlyHisAlaG lyHisHisHisHisHisHisHisHisHisHisHis 28 His7 Disks large-associated protein 3 Uniprot: 095886 Homo sapiens 214-233 25 N / A ProGlySerGlyGlyProHisThrSerHisHisHisHisHisHisH isHisHisHisHisHisGlnSerArgHis 29 His8 Nuclear receptor subfamily 4 group A member 3 Uniprot: Q92570 Homo sapiens 91-115 25 N / A AlaProSerTyrHisHisHisHisHisHisHisHisHisHisHisH isHisHisGlnGlnGlnHisGlnGlnPro 30 His9 E3 SUMO-protein ligase CBX4 Uniprot: 000257 Homo sapiens 377-400 24 N / A ThrHisProProSerHisHisProHisProHisProHisHisHisH isHisHisHisHisHisHisHisHis 31 HislO SKI / DACH domaincontaining protein 1 Uniprot: Q1XH10 Homo sapiens 334-371 38 N / A ProProProHisHisHisHisHisHisHisHisHisHisHisHisH isHisHisArgAlaGlnProProGlnGlnSerHisHisProProHi sHisHisArgProGlnProHis 32 Hisll GS homeobox 2 Uniprot: Q9BZM3 Homo sapiens 123-140 18 N / A AsnHisAlaHisHisHisHisHisProProGlnHisHisHisHisH isHisGln 33 Hisl2 N / A N / A N / A N / A 37 selfdesigned AlaGlyHisHisHisHisHisHisHisHisHisHisGlyGlyAlaG lyGlyAlaGlyGlyAlaGlyGlyAlaHisHisHisHisHisHisHi sHisHisHisGlyGlyAla 34 Hisl3 N / A N / A N / A N / A 37 selfdesigned AlaGlyHisHisHisHisHisHisHisHisHisHisSerSerGlyS erGlySerSerGlySerGlySerSerHisHisHisHisHisHisHi sHisHisHisGlyGlyAla 35 WO 2021 / 094746                                                 PCT / GB2020 / 052866 Hisl4 N / A N / A N / A N / A 17 self designed AlaGlyHisHisHisHisHisHisHisHisHisHisHisHisGlyG lyAla 36 Hisl5 N / A N / A N / A N / A 24 selfdesigned AlaGlyHisHisHisHisHisHisHisHisHisHisHisHisHisH isHisHisHisHis HisGlyGlyAla 37 super+ve single stranded DNA-binding protein GenBank: AWM11745.1 Escherichia phage vB_EcoM_NBG 1 256-299 74 selfdesigned: double sequence of MEAFSSAKT EDDFMSSSS SDDSDLDDL LAGL (SEQ ID NO: 131)and change amino acids D / E to K LysLysValAlaSerLysLeuLysPheLysLysLysMetGluAlaP heSerSerAlaLysThrLysLysLysPheMetSerSerSerSerSe rLysLysSerLysLeuLysLysleuLeuAlaGlyLeuMetGluAla PheSerSerAlaLysThrLysLysLysPheMetSerSerSerSerS erLysLysSerLysLeuLysLysLeuLeuAlaGlyLeu 38 super-ve single stranded DNA-binding protein GenBank: AWM11745.1 Escherichia phage vB_EcoM_NBG 1 256-299 75 selfdesigned: double sequence of MEAFSSAKT EDDFMSSSS SDDSDLDDL LAGL (SEQ ID NO: 131)and change amino acid K to D / E AspAspValAlaSerAspLeuAspAspPheAspAspAspMetGluA laPheSerSerAlaAspThrGluAspAspPheMetSerSerSerSe rSerAspAspSerAspLeuAspAspLeuLeuAlaGlyLeuMetGlu AlaPheSerSerAlaAspThrGluAspAspPheMetSerSerSerS erSerAspAspSerAspLeuAspAspLeuLeuAlaGlyLeu 39 superpositive RecA-like recombination protein GenBank: AYP69747.1 Escherichia phage vB EcoM Dal Ca~ 348-392 90 selfdesigned: double sequence ArglleValLysAlaLysValLysArgLeuIleAsnSerLysValL ysLysPheLysSerProLysSerLysSerLysSerAlaAlaLysLe uLysThrLysLeuLysGlnLeuSerLysMetLysLysPheAsnLys IleValLysAlaLysValLysLysLeuIleAsnSerLysValArgL 40 WO 2021 / 094746                                                 PCT / GB2020 / 052866 of EIVEAEVDE LINSKVEKF KSPESKSKS AADLETDLE QLSDMEEFN SEQ ID NO: 132) and change amino acids D / E to R / K ysPheLysSerProLysSerLysSerLysSerAlaAlaLysLeuLy sThrLysLeuLysGlnLeuSerLysMetLysGluPheAsn oe supernegative RecA-like recombination protein GenBank: AYP69747.1 Escherichia phage vB EcoM Dal Ca~ 348-392 90 selfdesigned: double sequence of EIVEAEVDE LINSKVEKF KSPESKSKS AADLETDLE QLSDMEEFN SEQ ID NO: 132) and change amino acids R / K to D / E GluIleValGluAlaGluValAspGluLeuIleAsnSerGluValG luGluPheAspSerProGluSerAspSerAspGluAlaAlaAspLe uGluThrAspLeuGluGlnLeuSerAspMetGluGluPheAsnGlu IleValGluAlaGluValAspGluLeuIleAsnSerGluValGluA spPheAspSerFroGluSerAspSerAspGluAlaAlaAspLeuGl uThrAspLeuGluGlnLeuSerAspMetGluGluPheAsn 41 DoubleX RecA-like recombination protein GenBank: AYP69747.1 Escherichia phage vB EcoM Dal Ca~ 348-392 90 selfdesigned: a linker of FEF and double sequence of EIVEAEVDE LINSKVEKF KSPESKSKS AADLETDLE QLSDMEEFN GluIleValGluAlaGluValAspGluLeuIleAsnSerLysValG luLysPheLysSerProGluSerLysSerLysSerAlaAlaAspLe uGluThrAspLeuGluGlnLeuSerAspMetGluGluPheAsnGlu IleValGluAlaGluValAspGluLeuIleAsnSerLysValGluL ysPheLysSerProGluSerLysSerLysSerAlaAlaAspLeuGl uThrAspLeuGluGlnLeuSerAspMetGluGluPheAsn 42 WO 2021 / 094746                                                 PCT / GB2020 / 052866 Triplextail RecA-like recombination protein GenBank: AYP69747.1 Escherichia phage vB EcoM Dal Ca- 348-392 SEQ ID NO: 132) selfdesigned: a linker of DDVASEF (SEQ ID NO: 133) and triple sequence of EIVEAEVDE LINSKVEKF KSPESKSKS AADLETDLE QLSDMEEFN (SEQ ID NO: 132) GluIleValGluAlaGluValAspGluLeuIleAsnSerLysValG luLysPheLysSerProGluSerLysSerLysSerAlaAlaAspLe uGluThrAspLeuGluGlnLeuSerAspMetGluGluPheAsnGlu IleValGluAlaGluValAspGluLeuIleAsnSerLysValGluL ysPheLysSerProGluSerLysSerLysSerAlaAlaAspLeuGl uThrAspLeuGluGlnleuSerAspMetGluGluPheAsnGluIle ValGluAlaGluValAspGluLeulleAsnSerLysValGluLysP heLysSerProGluSerLysSerLysSerAlaAlaAspLeuGluTh rAspLeuGluGlnLeuSerAspMetGluGluPheAsn 43 WO 2021 / 094746                                                 PCT / GB2020 / 052866 2026234101   18 Sep 2026 Like-for-like substitutions may be made, in the case of amino acids such as basic for basic, acidic for acidic, polar for polar, etc. Non-homologous substitution may also occur, i.e. from one class of residue to another, or alternatively involving the inclusion of unnatural amino acids such as ornithine, diaminobutyric acid ornithine, norleucine 5 ornithine, pyriylalanine, thienylalanine, naphthylalanine and phenylglycine. The specific IDR amino acid sequences disclosed herein (see Table 1) can broadly be classified into four groups. Some IDR sequences can be classified into more than one group. The RGG / RG group includes IDR sequences which are FG / YG rich. This group includes fib, hnrpnAl, DDX, HRP1 and Sup. The Poly Q group includes 10 IDR sequences which are Q / N rich. This group includes PCF11, Ent-1, HRP1, Sup, His4, His8 and HislO. The Poly P group includes sequences which are P rich. This group includes His4, His9 and HislO. The Poly H group includes sequences which are H rich. This group includes Hisl-11. Some key features of the IDR amino acid sequences are that they demonstrate cation-pi interactions and pi-pi interactions, and 15 are capable of forming amide bridges and salt bridges. Key features and key inter / intra molecular interactions of preferred IDR amino acid sequences are presented in Tables 2 to 20 below. Table 2 IDR fib Sequence PGFSPRGGGFGGRGGFGDRGGRGGRGGFGGGRGRGGGFRGRGR (SEQIDNO: 1) Key feature RGG / RG, FG Key inter / intra molecular interactions R-F / H (cation-pi interaction) F / H-F / H (pi-pi interaction) R-D (salt bridge) 2026234101   18 Sep 2026 Table 3 IDR HNRPNA1 ASASSSQRGRSGSGNFGGGRGGGFGGNDNFGRGGNFSGRGGFGGS Sequence RGGGGYGGSGDGYNGFGNDGSNFGGGGSYNDFGNYNNQSSNFGP MKGGNFGGRSSGPYGGGGOYFAKPONOGGYGVSSSSSSYGSGRRF (SEQIDNO: 12) Key feature RGG / RG, FG / YG Key R / K-Y / F / H (cation-pi interaction) inter / intra Y / F / H-Y / F / H (pi-pi interaction) molecular R / K-D (salt bridge) interactions Q / N-Q / N (amide interaction) Table 4 IDR DDX Sequence MGDEDWEAEINPHMSSYVPIFEKDRYSGENGDNFNRTPASSSEMDD GPSRRDHFMKSGFASGRNFGNRDAGECNKRDNTSTMGGFGVGKSF GNRGFSNSRFEDGDSSGFWRESSNDCEDNPTRNRGFSKRGGYRDGN NSEASGPYRRGGRGSFRGCRGGFGLGSPNNDLDPDECMORTGGLFG SRRPVLSGTGNGDTSQSRSGSGSERGGYKGLNEEVITGSGKNSWKS EAEGGESSDTQ (SEQ ID NO: 11) Key feature RGG / RG, FG Key inter / intra R / K-Y / F / H (cation-pi interaction) Y / F / H-Y / F / H (pi-pi interaction) R / K-D / E (salt bridge) 2026234101   18 Sep 2026 molecular Q / N-Q / N (amide interaction) interactions Table 5 IDR HRP1 Sequence GGNNGGNNMNRRGGNFGNQGDFNQMYQNPMMGGYNPMMNPQA MTDYYQKMQEYYOQMQ (SEQ ID NO: 9) Key feature RGG / FG, Q / N rich, YYQ-K / Q-MQ repeat Key inter / intra Q / N-Q / N(amide bridge) R / K-Y / F / H (cation-pi interaction) molecular Y / F / H-Y / F / H (pi-pi interaction) interactions R / K-D / E (salt bridge) Table 6 IDR Sup MSDSNQGNNQQNYQQYSQNGNQQQGNNRYQGYQAYNAQAQPAG Sequence GYYONYOGYSGYOOGGYOQYQYNPOGGYOOYNPOGGYOOYNPO GGYQQOFNPQGGRGNYKNFNYNNNLOGYQ (SEQ ID NO: 18) Key feature YNPQGGYQQ (SEQ ID NO: 19) repeats, RG, Q / N rich Key Q / N-Q / N (amide bridge) inter / intra R / K-Y / F / H (cation-pi interaction) molecular Y / F / H-Y / F / H (pi-pi interaction) interactions R / K-D (salt bridge) 2026234101   18 Sep 2026 Table 7 IDR PolCTD SPFGAYGEAPTSPGFGVSSPGFSPTSPTYSPTSPAYSPTSPSYSPTSPSY SPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPT Sequence SPSYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTSPAYSPTSPSYSPTSPS YSPTSPSYSPTSPSYSPTSPNYSPTSPSYSPTSPGYSPGSPAYSPKQDEQ (SEQIDNO: 13) Key repeats of YSPTSPS (SEQ ID NO: 124) and its variants (FSPTSPT (SEQ ID NO: 125), YSPTSP-A / N / G (SEQ ID NO: 126), YSPGSPA (SEQ ID feature NO: 127)) Key Q / N-Q / N(amide bridge) inter / intra K-Y / F / H (cation-pi interaction) molecular Y / F / H-Y / F / H (pi-pi interaction) interactions K-D / E (salt bridge) Table 8 IDR PCF11 Sequence QVQMOLROVFSQDOOVLQERMRYHELQOQQQQQYHETKDMVGS YTQNSNSAIPLFGNNSDTTNQQNS (SEQ ID NO: 7) Key feature Poly Q Key Q / N-Q / N(amide bridge) inter / intra R / K-Y / F / H (cation-pi interaction) molecular Y / F / H-Y / F / H (pi-pi interaction) interactions R / K-D / E (salt bridge) 2026234101   18 Sep 2026 Table 9 IDR ENT1 Sequence QNPTGISYSQPOQQQOPOOOPOYMONFOQQQPQYAONFOOQPQY TONYOOOPOYIOPHO (SEQ ID NO: 8) Key feature Poly Q, QQQPQY repeat (SEQ ID NO: 128) Key inter / intra molecular interactions Q / N-Q / N (amide bridge) Y / F / H-Y / F / H (pi-pi interaction) Table 10 IDR HIS1 Sequence PQQNALHHHHGNSSHHHHHHHHHHHHHGQQA (SEQ ID NO: 23) Key feature PolyH Key inter / intra molecular interactions Q / N-Q / N (amide bridge) H-H (pi-pi interaction) 2026234101   18 Sep 2026 Table 11 IDR HIS2 Sequence AGHHHHHPHAHHPLSQSSGHHHHHHHHHHQGYGGSG (SEQ ID NO: 24) Key feature PolyH Key inter / intra molecular interactions Q-Q (amide bridge) Y / H-Y / H (pi-pi interaction) Table 12 IDR HIS3 Sequence AGHHHHHHHHLPHLPPPHLHHHHHPQHHLHPGSAAAVHPVQQH (SEQ ID NO: 25) Key feature PolyH Key inter / intra molecular interactions Q-Q (amide bridge) H-H (pi-pi interaction) 2026234101   18 Sep 2026 Table 13 IDR HIS4 Sequence NHHASHGHHNSHHPQHHHHHHHHHHHPPPPAPQPPPPPQQQQ (SEQ ID NO: 26) Key feature Poly H, poly Q, poly proline Key inter / intra molecular interactions Q / N-Q / N (amide bridge) H-H (pi-pi interaction) Table 14 IDR HIS5 Sequence SGHHGAHHGAHHPAAAAAYEAFRGPGFAGGGGADDMGAGHHH GAHHAAHHHHAAHHHHHHHHHHGGAGHGGG AGHH (SEQ ID NO: 27) Key feature PolyH Key inter / intra molecular interactions R-Y / F / H(cation-pi interaction) Y / F / H-Y / F / H (pi-pi interaction) R-D / E (salt bridge) 2026234101   18 Sep 2026 Table 15 IDR HIS6 Sequence GGGGDHGGGGGHGHAGHHHHHHHHHHH (SEQ ID NO: 28) Key feature PolyH Key inter / intra molecular interactions H-H (pi-pi interaction) Table 16 IDR HIS7 Sequence PGSGGPHTSHHHHHHHHHHHHQSRH (SEQ ID NO: 29) Key feature PolyH Key inter / intra molecular interactions H-H (pi-pi interaction) R-H(cation-pi interaction) Q-Q (amide bridge) R-D / E (salt bridge) 2026234101   18 Sep 2026 Table 17 IDR HIS8 Sequence APSYHHHHHHHHHHHHHHQQQHQQP (SEQ ID NO: 30) Key feature Poly H, poly Q Key inter / intra molecular interactions Y / H-Y / H (pi-pi interaction) Q-Q (amide bridge) Table 18 IDR HIS9 Sequence THPPSHHPHPHPHHHHHHHHHHHH (SEQ ID NO: 31) Key feature Poly H, P rich Key inter / intra molecular interactions H-H (pi-pi interaction) 2026234101   18 Sep 2026 Table 19 IDR HIS10 Sequence PPPHHHHHHHHHHHHHHHRAQPPQQSHHPPHHHRPQPH (SEQ ID NO: 32) Key feature Poly H, P and Q rich Key inter / intra molecular interactions H-H (pi-pi interaction) Q-Q (amide bridge) R-H (cation-pi interaction) Table 20 IDR HIS11 Sequence NHAHHHHHPPQHHHHHHQ (SEQ ID NO: 33) Key feature PolyH Key inter / intra molecular interactions H-H (pi-pi interaction) Q / N-Q / N (amide bridge) 5   Functional properties of an I DR-macro molecule or IDR-polypeptide An IDR-tagged macromolecule or polypeptide, or IDR-macromolecule or IDR-polypeptide, as defined herein, must possess an amino acid sequence consisting of or 2026234101   18 Sep 2026 comprising one or more functional IDRs to be of use in the processes of the invention. Whether or not such an IDR amino acid sequence or domain is functional can be established by routine methods, such as those described herein. 5 Particle formation The inventors have surprisingly discovered that IDR-tagged polypeptides or IDR-polypeptides are capable of forming particles in an appropriate solution. This is believed to occur by liquid-liquid demixing leading to phase separation of fluids within 10 the solution mixture mediated by the IDR amino acid sequence. The formation of particles mediated by IDR amino acid sequences is described further in the examples below. The particles demonstrate a spheroidal appearance and can be described as “globules”, “globular foci” or “particles”. The terms “particle”, “globules” or “globular foci” as referred to herein are 15   intended to be synonymous and can be used interchangeably. Conditions and methods which permit the observation and detection of particles are set out herein including in the examples below. In the examples described herein, particle formation was observed to occur in a simple system comprising merely a solution of IDR-tagged polypeptide and divalent 20 metal cation. Particle formation was also found to occur in more complex mixtures including those comprising components required for RPA where one of the RPA protein components (Gp32) was IDR-tagged. In these situations, reaction components were seen to strongly co-localise with the particles, for example particles were seen to be dense in oligonucleotide as detected by the fluorescent labelled probe attached thereto, 25 as well comprising all other RPA reaction protein components. Detection and monitoring of the particles can be performed using any suitable method, as well as the methods set out in the examples below. Exemplary methods include microscopy, light scattering, flow cytometry, and microfluidic methods. The particles can be detected using microscopy, e.g., differential interference 30 contrast or fluorescence microscopy, to directly observe the particles at high magnification. With the aid of a computer, microscope images can be automatically 2026234101   18 Sep 2026 obtained and analyzed. Additionally, microscopy can allow for continual or frequent monitoring of at least a portion of a mixture containing particles. The particles can be detected using flow cytometry. In flow cytometry, one or more beams of light, e.g., each of a single wavelength, are directed onto a 5 hydrodynamically-focused stream of fluid. Suspended particles passing through the beams scatter the light, and fluorescent chemicals found in the particles or attached to the particles may be excited. The scattered and / or fluorescent light is analysed by detectors within the device, from which information about particle size and fluorescence can be determined. Modem flow cytometers can analyze several thousand particles 10 every second, in "real time", and can actively separate and isolate particles having specified properties. The particles can be detected using cytometry methods, devices, and systems as disclosed e.g. in US patent application publication nos. US2009 / 0079963 and US2010 / 0179068, and international patent application publication no. WO2009 / 112594. 15          The particles can be detected using microfluidic methods, devices, and systems. For example, the particles can be detected using a lab-on-a-chip device or system, or the like (see, e.g., US patent application publication nos. US2009 / 0326903 and US2009 / 0297733). Particles may be about 0.5-20 pm in size, e.g., between about any two sizes 20 selected from 0.5, 1,1.5, 2, 2.5, 3,4, 5, 6, 7, 8, 9, 10, 12, 15, 18, and 20 pm (e.g., about 1-10 pm in size). The concentration of particles may be approximately 10 to 5000 particles / nl, e.g., between any two numbers of particles selected from 10, 20, 50,100, 200, 500, 1000, 2000, and 5000 particles per nl, maybe detected (e.g. about 100-500 particles per 25 nl). The concentration of particles may be approximately 200-400 particles per nanolitre. Such phase-separated particles may be smaller than about 0.5 pm in size. Phase-separated particles, including those that are smaller than about 0.5 pm in size, may be detected by changes in the turbidity of the solution. Changes in the turbidity of 30 solutions can be measured by standard means and can be quantified typically according to the Formazin Turbidity Unit (FTU) or Formazin Nephelometric Unit (FNU). Other 2026234101   18 Sep 2026 methods include size exclusion chromatography, including multiangle light scattering (SEC-MALS). Experimental determination of IDR function 5 An IDR-macromolecule or IDR-polypeptide, or an IDR-tagged macromolecule or IDR-tagged polypeptide, as defined herein, can be determined to possess a functional intrincicahy disordered region (IDR) amino acid sequence and / or domain thereof, and thus to be of use in the processes and reagents of the invention, by using e.g. a phase 10   separation assay method or a RPA assay method as described below. Accordingly, an IDR-macromolecule, IDR-polypeptide or an IDR-tagged macromolecule or IDR-tagged polypeptide is a macromolecule or polypeptide which comprises, or which is tagged with, an amino acid sequence consisting of one or more functional intrinsically disordered regions; or a macromolecule or polypeptide which is 15   tagged with an amino acid sequence comprising one or more functional intrinsically disordered regions.. In all cases the functional intrinsically disordered region is one which may be determined to be functional in the phase separation assay method described below and / or in the RPA assay method as described below. 20 Phase separation assay method The phase separation assay method is a method comprising: 1. tagging the one or more intrinsically disordered region amino acid sequences to a polypeptide to create an IDR-polypeptide fusion protein, preferably tagged to 25         the recombinant phage vB EcoM NBG1 Gp32 protein to create a Gp32-IDR fusion protein, and providing purified IDR-polypeptide fusion protein; 2. adding the IDR-polypeptide fusion protein to a volume of water to a final concentration of 1000 ng / pl, wherein the final volume of the mixture is 50 pl, and preferably adding a divalent metal cation to a final concentration of 2 mM 30          or more, more preferably wherein the divalent metal cation is Mg2+, Mn2+, Ca2+, Co2+ orNi2+; 2026234101   18 Sep 2026 3. vortexing the mixure followed by pulse centrifuging the mixture; 4. transferring al 0 pl sample of the mixture supernatant to a haemocytometer slide; 5. viewing the haemocytometer slide under a microscope at x400 magnification; 5       6. observing the formation of particles in the mixture; 7. in the absence of particles in the mixture, repeating steps 1 to 6 and incrementally increasing the concentration of the divalent metal cation until the formation of particles in the mixture is observed; 8. counting the number of particles which form in a magnification area of 218 pm 10          x 175 pm at x400 magnification; and 9. (i) establishing that the amino acid sequence consisting of or comprising one or more intrinsically disordered regions (IDRs) is functional when 10 or more particles are counted in the magnification area, preferably when 50 or more particles are counted in the magnification area, more preferably when 100 or 15           more particles are counted in the magnification area; or (ii) establishing that the amino acid sequence consisting of or comprising one or more intrinsically disordered regions (IDRs) is non-functional if the concentration of the divalent metal cation is increased to 100 mM or more and no particles are observed to form in the magnification area, or less than 10 20           particles are counted in the magnification area. In the above method, should it be desirable to examine the effect of providing a divalent metal cation on particle formation, step 2 may comprise adding a divalent metal cation to any desired final concentration. The effects of different concentrations of divalent metal cations may thus be examined. 25          In the above method, should it be desirable to examine the effect of providing ATP on particle formation, step 2 may comprise adding ATP to any desired final concentration. The effects of different concentrations of ATP may thus be examined. ATP may be provided for example at a concentration of ImM to 3.5mM, e.g. ImM to 2mM. 30          Step 2 may comprise adding a detectable nucleic acid molecule and wherein step 8 comprises counting the number of particles by the detection means. For example, step 2026234101   18 Sep 2026 2 may comprise adding a probe having the nucleic acid sequence set for the in SEQ ID NO: 104, which is labelled with FAM (fluorescein) and step 8 may comprise detecting particles by fluorescence. The detectable nucleic acid molecule may be added to any suitable final concentration, such as 0.5pM. 5          Accordingly, the above assay may be used to examine reaction efficiency, the capability of causing liquid-liquid demixing and the capability of causing molecules to co-localise within a plurality of phase-separated aqueous compartments (particles). In the above method, if the divalent metal cation is Mg2+ the source of the cation is preferably MgOAc. If the divalent metal cation is Ca2+, the source of the cation is 10 preferably CaCh. If the divalent metal cation is Mn2+, the source of the cation is preferably MnCh. RPA assay method 15 20 25 The RPA assay method is a method comprising: 1. tagging the one or more intrinsically disordered region polypeptide sequences to a Gp32 protein, preferably the recombinant phage vB EcoM NBG1 Gp32 protein, to create a Gp32-IDR fusion protein and providing purified Gp32-IDR fusion protein; 2. creating a reaction mixture comprising a. Tris HC1 pH 8.3, 25 mM; b. KOAc, 7.5mM; c. DTT, 1 mM; d. ATP, 2.5 mM; e. phosphocreatine, 20 mM; f. creatine kinase, 1 pM; g. dNTPs, 1 mM; h. purified Gp32-IDR fusion protein, 20 pM; i. purified UvsX, 4.8 pM j. purified UvsY, 8.6 pM k. S. aureus DNA polymerase 1 (Sau), 0.135 pM 2026234101   18 Sep 2026 1. Exonuclease III, 0.27 pM m. forward primer, 0.4 pM n. reverse primer, 0.4 pM o. probe, 0.12 pM 5       3. initiating a recombinase polymerase amplification reaction by adding to the reaction mixture 33 mM MgOAc and 10 copies of template nucleic acid; 4. incubating the reaction mixture at 39°C in a fluorometer with magnetic mixing using a bearing ball; 5. (i) establishing that the one or more intrinsically disordered region polypeptide 10          sequence is functional when a 2-fold or more more increase in amplified product is detectable within 15 minutes by a measurable increase in fluorescence compared to baseline in a template-dependent fashion; or (ii) establishing that the one or more intrinsically disordered region polypeptide sequence is non-functional when no amplified product is detected by 15           fluorescence after 15 minutes. In the above method, the forward primer sequence is: CGCCTGCAAGTCCTAAGACGCCAATCGAAAAGAAAC (SEQ ID NO:98); the reverse primer sequence is: CTGCATCTCCGTGGTATACTAATACATTGTTTTTA (SEQ ID NO:99); the probe sequence is: 20 CGAAAAGAAACACGCGGATGAAATCGATAAG[FAM] [THF][BHQ-1]ATACAAGGATTGGA (SEQ ID NO: 100), where FAM is fluorescein, THF is tetrahydrofuran and BHQ is Black Hole Quencher; and the template is Listeria monocytogenes genomic DNA. Step 5 of the above RPA assay method may alternatively comprise establishing 25 that the one or more intrinsically disordered region polypeptide sequence is functional when a 5-fold or more more increase in amplified product is detectable within 15 minutes by a measurable increase in fluorescence compared to baseline in a templatedependent fashion, or when a 10-fold or more more increase is detectable, or a 20-fold or more more increase, or a 30-fold or more more increase, or a 40-fold or more more 30 increase, or a 50-fold or more more increase, or a 100-fold or more more increase, or a 150-fold or more more increase, or a 200-fold or more more increase, or a 250-fold or 2026234101   18 Sep 2026 more more increase, or a 300-fold or more more increase, or a 350-fold or more more increase, or a 400-fold or more more increase, or a 450-fold or more more increase, or a 500-fold or more, 1000-fold or more, 2000-fold or more, 3000-fold or more, 4000-fold or more or 5000-fold or more more increase. By an increase in amplified product over 5 baseline it is meant the increase in amplified product compared to the amount of amplified product obtained by performing the reaction under the same conditions except wherein the Gp32 protein has not been tagged with the one or more intrinsically disordered region polypeptide sequences. In the above method, should it be desirable to examine the effect of providing a 10   divalent metal cation on reaction efficiency, step 3 may comprise adding a divalent metal cation to any desired final concentration. The effects of different concentrations of divalent metal cations may thus be examined. In the above method, should it be desirable to examine the effect of providing ATP on reaction efficiency, step 2 may comprise adding ATP to any desired final 15   concentration. The effects of different concentrations of ATP may thus be examined. ATP may be provided for example at a concentration of ImM to 3.5mM, e.g. ImM to 2mM. Tagging of IDR amino acid sequences to macromolecule and polypeptides 20 The methods, processes and reagents of the present invention involve, inter alia, IDR-tagged macromolecules and IDR-tagged polypeptides, wherein an IDR-tagged macromolecule or an IDR-tagged polypeptide is a macromolecule or polypeptide of interest tagged with an amino acid sequence consisting of or comprising one or more 25 intrinsically disordered regions (IDRs) (which may be referred to herein as an IDR moiety). The term “tag” or “tagging” is to be understood in its broadest sense. The terms are to be understood to mean that an IDR moiety, i.e. an amino acid sequence which consists of or comprises one or more functional IDRs, is attached to, tethered to, bound 30 to or otherwise associated with a macromolecule or a polypeptide of interest in any suitable way. 2026234101   18 Sep 2026 The most preferred means by which an IDR moiety is tagged to a polypeptide of interest is by creating a recombinant genetic fusion protein, wherein the polypeptide of interest is genetically engineered at the nucleotide level such that when transcribed and translated the expressed protein comprises the polypeptide of interest together with the 5 IDR moiety. If desired, linkers may be placed between the polypeptide of interest and the IDR moiety. For example, flexible, ridgid and cleavable linkers are well known in the art and are widely used in the manufacture of fusion proteins (see, e.g.: Fusion Protein Linkers: Property, Design and Functionality, Chen, X., et al. 2013, Adv. Drug Deliv. 10 Rev., 15, 65(10), ppl357-1369). Standard methods for genetic engineering are well known in the art (see for example, Sambrook et al., 2001, Molecular Cloning: a Laboratory Manual, 3rd edition, Cold Spring Harbour Laboratory Press; and Current Protocols in Molecular Biology, Greene Publishing and Wiley-lnterscience, New York (1995)), as are methods for 15   protein expression and purification. Other means by which an IDR moiety may be tagged to a macromolecule or polypeptide of interest is by one or more covalent bonds or by an affinity interaction. The IDR moiety can be tagged to a polypeptide in any suitable orientation, such as at the N-terminus of the polypeptide of interest, at the C-terminus of the polypeptide 20   of interest, or the polypeptide of interest may comprise an IDR moiety at both its N- terminus and C-terminus, or at any amino acid position along the length of the polypeptide. Peptides / oligopeptides / polypeptides / proteins may be attached / tethered conjugated to other macromolecules, incuding other 25 peptides / oligopeptides / polypeptides / proteins, by the use of methods very known in this technical field. One such method is “click chemistry”. The term “click chemistry” is typicaly used to describe the reaction of an azide with an alkyne resulting in a 1,5-disubstituted 1,2,3-triazole in the presence of a copper catalyst. Click chemistry allows 30 peptides / oligopeptides / polypeptides / proteins to be conjugated to a vast range of other macromolecules including other peptides / oligopeptides / polypeptides / proteins, as well as 2026234101   18 Sep 2026 e.g. carbohydrates, nucleic acids, polymers, drugs, aptamers, hydrogels etc. The method is also referred to as “CuAAC” (Cu catalyzed alkyne azide cycloaddition) (see e.g. “Click” reactions: a versatile toolbox for the synthesis of peptide-conjugates. Tang, W. etal., 2014, Chern. Soc. Rev.,43, pp7013-7039). 5           Many other linker / crosslinker chemistries are available to conjugate peptides / oligopeptides / polypeptides / proteins to other macromolecules, such as crosslinkers which contain maleimide, sulfhydryl reactive groups or succinimidyl esters (often referred to as NHS esters), which react with amines. For example, succinimides can be used to form covalent bonds between proteins or peptides and plastics materials. 10          Standard chemistries can be used which are commonly employed to create conjugates between polypeptides and non-polypeptide molecules, such as chemistries to create antibody-drug conjugates. Many such techniques are well known in this technical field. Affinity-based interactions can also be employed. For example, an amino acid 15   sequence consisting of or comprising one or more functional intrinsically disordered regions may be attached / tethered to a macromolecule or polypeptide of interest by an affinity-based interaction, such as streptavidin-biotin, receptor-ligand interaction etc. Multivalent metal cations for IDR amino acid sequence function 20 When IDR-macromolecules or IDR-polypeptides, as described and defined herein, are used in in vitro biochemical reactions, the in vitro biochemical reaction buffer preferably contains multivalent metal cations, preferably divalent metal cations. The presence of multivalent / divalent metal cations in the reaction buffer helps to 25 facilitate and enhance liquid-liquid demixing leading to phase separation in the in vitro biochemical reaction mileiu mediated / caused by an IDR-macromolecule or an IDR-polypeptide. The functional capability of a divalent metal cation to enhance phase separation in an in vitro biochemical reaction mileiu mediated / caused by an IDR-macromolecule 30 or IDR-polypeptide can readily be established, such as by techniques disclosed and defined herein. In particular, such functional capability can be established by the ability 2026234101   18 Sep 2026 of the multivalent / divalent metal cation to induce the formation of globular foci or particles, as further described and defined herein, in the in vitro biochemical reaction mileiu in an IDR-dependent manner, for example as determined by assays described herein. 5          The use of a divalent metal cation in promoting / enhancing IDR-dependent liquid-liquid demixing leading to phase separation is preferred. However, functional equivalents of any multivalent or any divalent metal cation are envisaged. A functional equivalent of a multivalent / divalent metal cation as described herein is any agent that may substitute for a divalent metal cation in promoting IDR-dependent liquid-liquid 10   demixing leading to phase separation in an in vitro biochemical reaction milieu, for example as determined by assays described herein. Any suitable multivalent / divalent metal cation may be used, either as a single agent or combination of agents and optionally in the presence of a chelating agent such as ethylenediaminetetraacetic acid (EDTA), ethylene glycol-bis(P-aminoethyl ether)- 15 N,N,N',N'-tetraacetic acid (EGTA) or nitriloacetic acid (NTA). The divalent metal cation may be Mg2+, Mn2+, Ca2+, Co2+, Ni2+ or Cu2+. Any of these cations may be used as a single agent, or any combination of cations may be used. Preferably they are used as single agents. Preferred divalent metal cations are Mg2+, Mn2+ and Ca2+. 20          The specific multivalent / divalent metal cation which achieves optimal results in promoting IDR-mediated phase separation in an in vitro biochemical reaction milieu, as well as the specific concentration of the multivalent / divalent metal cation used may depend upon the specific intrinsically disordered region amino acid sequence which is used to tag the macromolecule or polypeptide of interest. The optimal 25 multivalent / divalent metal cation and optimal concentration can be established empirically using routine tests. Phase separation assays as described further herein may be used for this purpose. Preferred concentrations ranges of the multivalent / divalent metal cations are from about 300 pM to about 100 mM, from about 300 pM to about 50 mM, from about 30   400 pM to about 50 mM, from about 400 pM to about 20 mM, from about 400 pM to 2026234101   18 Sep 2026 about 30 mM, from about 500 pM to about 10 mM, from about 500 pM to about 25 mM and from about 1 mM to about 35 mM. An in vitro biochemical reaction buffer may contain Mg2+ ions. A preferred concentration range is from about 300 pM to about 100 mM, more preferably from 5 about 400 pM to about 50 mM, yet more preferably from about 500 pM to about 40 mM, even more preferably from about 25 mM to about 35 mM, such as 33 mM . Preferably the buffer contains MgOAc at the indicated concentrations. An in vitro biochemical reaction buffer may contain Ca2+ ions. A preferred concentration range is from about 300 pM to about 100 mM, more preferably from 10 about 400 pM to about 50 mM, yet more preferably from about 1 mM to about 40 mM, even more preferably from about 25 mM to about 35 mM, such as 33 mM . Preferably the buffer contains CaCh at the indicated concentrations. An in vitro biochemical reaction buffer buffer may contain Mn2+ ions. A preferred concentration range is from about 300 pM to about 50 mM, more preferably 15 from about 400 pM to about 50 mM, yet more preferably from about 500 pM to about 40 mM, even more preferably from about 25 mM to about 35 mM, such as 33 mM. Preferably the buffer contains MnCh at the indicated concentrations. Recombinase polymerase amplification (RPA) 20 Recombinase polymerase amplification (RPA) is a method for isothermal amplification of nucleic acids. In general, in a first step of RPA, a recombinase agent is contacted with first and second nucleic acid primers and a recombinase loading protein to form first and second nucleoprotein primers. In general, in a second step, the first 25 and second nucleoprotein primers are contacted with a double stranded template nucleic acid to form a first double stranded structure at a first portion of the first strand of the template nucleic acid, and a second double stranded structure at a second portion of the second strand of the template nucleic acid such that the 3 ’ ends of the first nucleic acid primer and the second nucleic acid primer are orientated towards each other on a given 30 nucleic acid molecule. In general, in a third step, the 3’ end of the first and the second nucleoprotein primers are extended by polymerase to generate first and second double 2026234101   18 Sep 2026 stranded nucleic acids, and first and second displaced single strands of nucleic acid. A single stranded stabilizing agent is employed to stabilize the first and second displaced single strands of nucleic acid. Generally, the second and third steps can be repeated until a desired degree of amplification is reached. 5          RPA methods are disclosed extensively, e.g., in US Patent No. 7,270,981; US Patent No. 7,399,590; US Patent No. 7,666,598; US Patent No. 7,435,561 and International Patent Application Publication No. WO2010 / 141940. In addition, for a comprehensive recent review see: Review: a comprehensive summary of a decade development of the recombinase polymerase amplification, Li, J. et al., 2019, Analyst, 10    144, pp31-67). Recombinase agent RPA methods, including those of the present invention, use a recombinase 15    agent. Any of the one or more IDR-polypeptides of the present invention may be attached / tethered / tagged to any recombinase agent. A recombinase agent is a molecule, typically an enzyme, that can coat a single stranded nucleic acid, typically DNA (ssDNA) to form a nucleoprotein filament. Such 20 filaments can then “scan” a double stranded nucleic acid molecule, typically DNA (dsDNA) for regions of sequence homology / complementarity. When complementary sequences are located, the nucleoprotein filament (comprising the recombinase agent) strand invades the double stranded nucleic acid molecule creating a short hybrid and a displaced strand bubble known as a D-loop. 25         Any suitable recombinase agent may be used in the RPA methods described herein, and may be tagged with any of the IDR amino acid sequences as described herein. The recombinase agent may originate from a prokaryotic, eukaryotic or viral organism. 30         The recombinase agent may be RecA, UvsX, RadA, RadB, Rad 51 or any functional variant, analoge, homologe or derivative of any of these proteins. 2026234101   18 Sep 2026 Any combination of these proteins may be used. Suitable recombinase agents include the E. coli RecA protein, the T4 UvsX protein, or any homologous protein or protein complex from any phyla. Eukaryotic RecA homologues are generally named Rad51 after the first member 5   of this group to be identified. Other non-homologous recombinase agents may be utilized in place of RecA, for example RecT or RecO. Exemplary recombinase agents include RecA and UvsX, and fragments or mutants thereof and combinations thereof. The RecA and UvsX proteins can be obtained from any species. RecA and UvsX fragments or mutant proteins can also be 10 produced using the available RecA and UvsS protein and nucleic acids sequences, and molecular biology techniques. Exemplary UvsX proteins include those derived from myoviridae phages, such as T4, T2, T6, Rb69, Aehl, KVP40, Acinetobacter phage 133, Aeromonas phage 65, cyanophage P-SSM2, cyanophage PSSM4, cyanophage S-PM2, Rbl4, Rb32, Aeromonas phage 25, Vibrio phage nt-1, phi-1, Rbl6, Rb43, Phage 31, 15   phage 44RR2.81, Rb49, phage Rb3, and phage LZ2. Additional exemplary recombinase agents include archaebacterial RADA and RADB proteins and eukaryotic (e.g., plant, mammal, and fungal) Rad51 proteins (e.g., RAD51, RAD51B, RAD51C, RAD51D, DMC1, XRCC2, XRCC3, and recA). The recombinase agent is preferably UvsX, T4 UvsX, T6 UvsX, RBI8 UvsX, 20 E.coli phage wV7 UvsX, Shigella phage CB8 UvsX, Shigella phage Shfl2 UvsX, E.coli phage ARI UvsX, phage vB_EcoM_G4507 UvsX, Shigella phage SHFML-11 UvsX, Escherichia phage vB_EcoM_DalCa UvsX, E. coli RecA, E. coli RadA, E. coli RadB, E. coli Rad 51 or any functional variant, analog, homolog or derivative thereof or any combination thereof. A particularly preferred recombinase agent is Escherichia phage 25 vBEcoMDalCa UvsX. The recombinase agent may also comprise a C-terminal deletion of acidic residues to improve its activity. Any functional variants, analogs, homologs or derivatives of the recombinase agent above may also function itself as a recombinase agent and these functional 30 variants, analogs, homologs or derivatives are also contemplated as a recombinase agent to be used in the processes described and defined herein. 2026234101   18 Sep 2026 For example, a small peptide from RecA, has been shown to retain some aspects of the recombination properties of RecA. This peptide comprises residues 193 to 212 of E. coli RecA and can mediate pairing of single stranded oligonucleotides. The recombinase agent (e.g., UvsX) may be a mutant or hybrid recombinase 5 agent. Mutant forms of UvsX are described in U.S. Patent No. 8,071,308. The mutant UvsX may be an Rb69 UvsX that includes at least one mutation in the Rb69 UvsX amino acid sequence, wherein the mutation is selected from the group consisting of (a) an amino acid which is not histidine at position 64, a serine at position 64, the addition of one or more glutamic acid residues at the C-terminus, the addition of one or more 10 aspartic acid residues at the C-terminus, and a combination thereof. The mutant UvsX may be a T6 UvsX having at least one mutation in the T6 UvsX amino acid sequence, wherein the mutation is selected from the group consisting of (a) an amino acid which is not histidine at position 66; (b) a serine at position 66; (c) the addition of one or more glutamic acid residues at the C-terminus; (d) the addition of 15   one or more aspartic acid residues at the C-terminus; and (e) a combination thereof. Where a hybrid recombinase agent is used, the hybrid protein may, for example, be a UvsX protein that includes at least one region that includes an amino acid sequence derived from a different UvsX species. The region may be, for example, the DNA-binding loop-2 region of UvsX. 20          If desired, the recombinase agent may be a temperature-sensitive (referred to herein as "ts") recombinase agent. If a ts recombinase agent is used, the RPA reaction may be started at one temperature (the permissive temperature) and terminated at another temperature (the non-permissive temperature). Combinations of permissive temperatures maybe, for example 25°C / 30°C, 30°C / 37°C, 37°C / 42°C and the like. The 25 ts protein may be reversible. A reversible ts protein’s activity is restored when it is shifted from the non-permissive temperature to the permissive temperature. While any recombinase agent concentration may be used, preferred recombinase concentrations may be, for example, in the range of 0.2-12 pM, 6-12 pM, 4-12 pM and 4-6 pM, preferably about 5 pM, more preferably about 4.8 pM. 30          Recombinase agents generally require the presence of ATP, ATPyS, or other nucleoside triphosphates or their analogs. It is preferred that recombinase agents are 2026234101   18 Sep 2026 used in a reaction environment in which regeneration of targeting sites can occur shortly following a round of D-loop stimulated synthesis. Completed recombination events involving recombinase disassembly will avoid a stalling of amplification or very inefficient linear amplification of ssDNA caused by oscillating single sided synthesis 5 from one end to the other. Exemplary UvsX recombinase agents tagged with amino acid tag sequences comprising intrinsically disordered regions are set out in Table 21 below. Table 21 Name of protein SEQID NO. Amino acid sequence UvsX (7His) 44 MSIADLKSRLIKASTSKMTAELTTSKFFNEKDVIRTKIP MLNIAISGAIDGGMQSGLTIFAGPSKSFKSNMSLTMV AAYLNKYPDAICLFYDSEFGITPAYLKSMGVDPERVI HTPIQSVEQLKIDMVNQLETIERGEKVIVFIDSIGNMAS KKETEDALNEKSVADMTRAKSLKSLFRIVTPYFSIKNI PCVAVNHTIETIEMFSKTVMTGGTGVMYSADTVFIIG KRQIKDGSDLQGYQFVLNVEKSRTVKEKSKFFIDVKF DGGIDPYSGLLDMALELGFVVKPKNGWYAREFLDEE TGEMIREEKSWRAKDTNCTTFWGPLFKHQPFRDAIKR AYQLGAIDSNEIVEAEVDELINSKVEKFKSPESKSKSA ADLETDLEQLSDMEEFNEHHHHHHH UvsX-fib-1 45 MSIADLKSRLIKASTSKMTAELTTSKFFNEKDVIRTKIP MLNIAISGAIDGGMQSGLTIFAGPSKSFKSNMSLTMV AAYLNKYPDAICLFYDSEFGITPAYLKSMGVDPERVI HTPIQSVEQLKIDMVNQLETIERGEKVIVFIDSIGNMAS KKETEDALNEKSVADMTRAKSLKSLFRIVTPYFSIKNI PCVAVNHTIETIEMFSKTVMTGGTGVMYSADTVFIIG KRQIKDGSDLQGYQFVLNVEKSRTVKEKSKFFIDVKF 2026234101   18 Sep 2026 DGGIDPYSGLLDMALELGFVVKPKNGWYAREFLDEE TGEMIREEKSWRAKDTNCTTFWGPLFKHQPFRDAIKR AYQLGAIDSNEIVEAEVDELINSKVEKFKSPESKSKSA ADLETDLEQLSDMEEFNEPGFSPRGGGFGGRGGFGDR GGRGGRGGFGGGRGRGGGFRGRGRHHHHHHH UvsX-fib-2 46 MSIADLKSRLIKASTSKMTAELTTSKFFNEKDVIRTKIP MLNIAISGAIDGGMQSGLTIFAGPSKSFKSNMSLTMV AAYLNKYPDAICLFYDSEFGITPAYLKSMGVDPERVI HTPIQSVEQLKIDMVNQLETIERGEKVIVFIDSIGNMAS KKETEDALNEKSVADMTRAKSLKSLFRIVTPYFSIKNI PCVAVNHTIETIEMFSKTVMTGGTGVMYSADTVFIIG KRQIKDGSDLQGYQFVLNVEKSRTVKEKSKFFIDVKF DGGIDPYSGLLDMALELGFVVKPKNGWYAREFLDEE TGEMIREEKSWRAKDTNCTTFWGPLFKHQPFRDAIKR AYQLGAIDSNEIVEAEVDELINSKVEKFKSPESKSKSA ADLETDLEQLSDMEEFNEPGFSPRGGGFGGRGGFGDR GGRGGRGGFGGGRGRGGVEHHHHHH UvsX-fib-3 47 MSIADLKSRLIKASTSKMTAELTTSKFFNEKDVIRTKIP MLNIAISGAIDGGMQSGLTIFAGPSKSFKSNMSLTMV AAYLNKYPDAICLFYDSEFGITPAYLKSMGVDPERVI HTPIQSVEQLKIDMVNQLETIERGEKVIVFIDSIGNMAS KKETEDALNEKSVADMTRAKSLKSLFRIVTPYFSIKNI PCVAVNHTIETIEMFSKTVMTGGTGVMYSADTVFIIG KRQIKDGSDLQGYQFVLNVEKSRTVKEKSKFFIDVKF DGGIDPYSGLLDMALELGFVVKPKNGWYAREFLDEE TGEMIREEKSWRAKDTNCTTFWGPLFKHQPFRDAIKR AYQLGAIDSNEIVEAEVDELINSKVEKFKSPESKSKSA ADLETDLEQLSDMEEFNEPGFSPRGGGFGGRGGFGDR GGRGGRGGVEHHHHHH 2026234101   18 Sep 2026 UvsX-fib-4 48 MSIADLKSRLIKASTSKMTAELTTSKFFNEKDVIRTKIP MLNIAISGAIDGGMQSGLTIFAGPSKSFKSNMSLTMV AAYLNKYPDAICLFYDSEFGITPAYLKSMGVDPERVI HTPIQSVEQLKIDMVNQLETIERGEKVIVFIDSIGNMAS KKETEDALNEKSVADMTRAKSLKSLFRIVTPYFSIKNI PCVAVNHTIETIEMFSKTVMTGGTGVMYSADTVFIIG KRQIKDGSDLQGYQFVLNVEKSRTVKEKSKFFIDVKF DGGIDPYSGLLDMALELGFVVKPKNGWYAREFLDEE TGEMIREEKSWRAKDTNCTTFWGPLFKHQPFRDAIKR AYQLGAIDSNEIVEAEVDELINSKVEKFKSPESKSKSA ADLETDLEQLSDMEEFNEPGFSPRGGGFGGRGGVEH HHHHH UvsX- HNRNPA1 49 MSIADLKSRLIKASTSKMTAELTTSKFFNEKDVIRTKIP MLNIAISGAIDGGMQSGLTIFAGPSKSFKSNMSLTMV AAYLNKYPDAICLFYDSEFGITPAYLKSMGVDPERVI HTPIQSVEQLKIDMVNQLETIERGEKVIVFIDSIGNMAS KKETEDALNEKSVADMTRAKSLKSLFRIVTPYFSIKNI PCVAVNHTIETIEMFSKTVMTGGTGVMYSADTVFIIG KRQIKDGSDLQGYQFVLNVEKSRTVKEKSKFFIDVKF DGGIDPYSGLLDMALELGFVVKPKNGWYAREFLDEE TGEMIREEKSWRAKDTNCTTFWGPLFKHQPFRDAIKR AYQLGAIDSNEIVEAEVDELINSKVEKFKSPESKSKSA ADLETDLEQLSDMEEFNEASAS S SQRGRSGSGNFGGG RGGGFGGNDNFGRGGNFSGRGGFGGSRGGGGYGGS GDGYNGFGNDGSNFGGGGSYNDFGNYNNQSSNFGP MKGGNFGGRSSGPYGGGGQYFAKPQNQGGYGVSSSS SSYGSGRRFHHHHHHH 2026234101   18 Sep 2026 UvsX-DDX 50 MSIADLKSRLIKASTSKMTAELTTSKFFNEKDVIRTKIP MLNIAISGAIDGGMQSGLTIFAGPSKSFKSNMSLTMV AAYLNKYPDAICLFYDSEFGITPAYLKSMGVDPERVI HTPIQSVEQLKIDMVNQLETIERGEKVIVFIDSIGNMAS KKETEDALNEKSVADMTRAKSLKSLFRIVTPYFSIKNI PCVAVNHTIETIEMFSKTVMTGGTGVMYSADTVFIIG KRQIKDGSDLQGYQFVLNVEKSRTVKEKSKFFIDVKF DGGIDPYSGLLDMALELGFVVKPKNGWYAREFLDEE TGEMIREEKSWRAKDTNCTTFWGPLFKHQPFRDAIKR AYQLGAIDSNEIVEAEVDELINSKVEKFKSPESKSKSA ADLETDLEQLSDMEEFNEMGDEDWEAEINPHMSSYV PIFEKDRYSGENGDNFNRTPASSSEMDDGPSRRDHFM KSGFASGRNFGNRDAGECNKRDNTSTMGGFGVGKSF GNRGFSNSRFEDGDSSGFWRESSNDCEDNPTRNRGFS KRGGYRDGNNSEASGPYRRGGRGSFRGCRGGFGLGS PNNDLDPDECMQRTGGLFGSRRPVLSGTGNGDTSQS RSGSGSERGGYKGLNEEVITGSGKNSWKSEAEGGESS DTQHHHHHHH UvsX addPolCTD 51 MSIADLKSRLIKASTSKMTAELTTSKFFNEKDVIRTKIP MLNIAISGAIDGGMQSGLTIFAGPSKSFKSNMSLTMV AAYLNKYPDAICLFYDSEFGITPAYLKSMGVDPERVI HTPIQ SVEQLKIDMVNQLETIERGEKVIVFIDSIGNMAS KKETEDALNEKSVADMTRAKSLKSLFRIVTPYFSIKNI PCVAVNHTIETIEMFSKTVMTGGTGVMYSADTVFIIG KRQIKDGSDLQGYQFVLNVEKSRTVKEKSKFFIDVKF DGGIDPYSGLLDMALELGFVVKPKNGWYAREFLDEE TGEMIREEKSWRAKDTNCTTFWGPLFKHQPFRDAIKR AYQLGAIDSNEIVEAEVDELINSKVEKFKSPESKSKSA ADLETDLEQLSDMEEFNEHHHHHHHFEFSPFGAYGE APTSPGFGVSSPGFSPTSPTYSPTSPAYSPTSPSYSPTSP 2026234101   18 Sep 2026 SYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTS PSYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPT SPSYSPTSPAYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSP TSPNYSPTSPSYSPTSPGYSPGSPAYSPKQDEQTAALE HHHHHH UvsX-fiisPolII 52 MSIADLKSRLIKASTSKMTAELTTSKFFNEKDVIRTKIP MLNIAISGAIDGGMQSGLTIFAGPSKSFKSNMSLTMV AAYLNKYPDAICLFYDSEFGITPAYLKSMGVDPERVI HTPIQSVEQLKIDMVNQLETIERGEKVIVFIDSIGNMAS KKETEDALNEKSVADMTRAKSLKSLFRIVTPYFSIKNI PCVAVNHTIETIEMFSKTVMTGGTGVMYSADTVFIIG KRQIKDGSDLQGYQFVLNVEKSRTVKEKSKFFIDVKF DGGIDPYSGLLDMALELGFVVKPKNGWYAREFLDEE TGEMIREEKSWRAKDTNCTTFWGPLFKHQPFRDAIKR AYQLGAIDSNEIVEAEVDELINSKVEKFKSPESKSKSA ADLETDLEQLSDMEEFNETSPSYSPTSPSYSPTSPGYSP TSPAYSPTSPTYSPTSPTYSPHHHHHHH UvsX-PCFll 53 MSIADLKSRLIKASTSKMTAELTTSKFFNEKDVIRTKIP MLNIAISGAIDGGMQSGLTIFAGPSKSFKSNMSLTMV AAYLNKYPDAICLFYDSEFGITPAYLKSMGVDPERVI HTPIQSVEQLKIDMVNQLETIERGEKVIVFIDSIGNMAS KKETEDALNEKSVADMTRAKSLKSLFRIVTPYFSIKNI PCVAVNHTIETIEMFSKTVMTGGTGVMYSADTVFIIG KRQIKDGSDLQGYQFVLNVEKSRTVKEKSKFFIDVKF DGGIDPYSGLLDMALELGFVVKPKNGWYAREFLDEE TGEMIREEKSWRAKDTNCTTFWGPLFKHQPFRDAIKR AYQLGAIDSNEIVEAEVDELINSKVEKFKSPESKSKSA ADLETDLEQLSDMEEFNEQVQMQLRQVFSQDQQVLQ 2026234101   18 Sep 2026 ERMRYHELQQQQQQQYHETKDMVGSYTQNSNSAIPL FGNNSDTTNQQNSHHHHHHH UvsX-Sup 54 MSIADLKSRLIKASTSKMTAELTTSKFFNEKDVIRTKIP MLNIAISGAIDGGMQSGLTIFAGPSKSFKSNMSLTMV AAYLNKYPDAICLFYDSEFGITPAYLKSMGVDPERVI HTPIQSVEQLKIDMVNQLETIERGEKVIVFIDSIGNMAS KKETEDALNEKSVADMTRAKSLKSLFRIVTPYFSIKNI PCVAVNHTIETIEMFSKTVMTGGTGVMYSADTVFIIG KRQIKDGSDLQGYQFVLNVEKSRTVKEKSKFFIDVKF DGGIDPYSGLLDMALELGFVVKPKNGWYAREFLDEE TGEMIREEKSWRAKDTNCTTFWGPLFKHQPFRDAIKR AYQLGAIDSNEIVEAEVDELINSKVEKFKSPESKSKSA ADLETDLEQLSDMEEFNEMSDSNQGNNQQNYQQYSQ NGNQQQGNNRYQGYQAYNAQAQPAGGYYQNYQGY SGYQQGGYQQYQYNPQGGYQQYNPQGGYQQYNPQ GGYQQQFNPQGGRGNYKNFNYNNNLQGYQHHHHH HH UvsX- DoubleX 55 MSIADLKSRLIKASTSKMTAELTTSKFFNEKDVIRTKIP MLNIAISGAIDGGMQSGLTIFAGPSKSFKSNMSLTMV AAYLNKYPDAICLFYDSEFGITPAYLKSMGVDPERVI HTPIQSVEQLKIDMVNQLETIERGEKVIVFIDSIGNMAS KKETEDALNEKSVADMTRAKSLKSLFRIVTPYFSIKNI PCVAVNHTIETIEMFSKTVMTGGTGVMYSADTVFIIG KRQIKDGSDLQGYQFVLNVEKSRTVKEKSKFFIDVKF DGGIDPYSGLLDMALELGFVVKPKNGWYAREFLDEE TGEMIREEKSWRAKDTNCTTFWGPLFKHQPFRDAIKR AYQLGAIDSNEIVEAEVDELINSKVEKFKSPESKSKSA ADLETDLEQLSDMEEFNE HHHHHHHFEFEIVEAEVDELINSKVEKFKSPESKSKSA 2026234101   18 Sep 2026 ADLETDLEQLSDMEEFNEIVEAEVDELINSKVEKFKSP ESKSKSAADLETDLEQLSDMEEFNHHHHHH Superpositive 56 MSIADLKSRLIKASTSKMTAELTTSKFFNEKDVIRTKIP MLNIAISGAIDGGMQSGLTIFAGPSKSFKSNMSLTMV AAYLNKYPDAICLFYDSEFGITPAYLKSMGVDPERVI HTPIQSVEQLKIDMVNQLETIERGEKVIVFIDSIGNMAS KKETEDALNEKSVADMTRAKSLKSLFRIVTPYFSIKNI PCVAVNHTIETIEMFSKTVMTGGTGVMYSADTVFIIG KRQIKDGSDLQGYQFVLNVEKSRTVKEKSKFFIDVKF DGGIDPYSGLLDMALELGFVVKPKNGWYAREFLDEE TGEMIREEKSWRAKDTNCTTFWGPLFKHQPFRDAIKR AYQLGAIDSNRIVKAKVKRLINSKVKKFKSPKSKSKS AAKLKTKLKQLSKMKKFNKIVKAKVKKLINSKVRKF KSPKSKSKSAAKLKTKLKQLSKMKEFNHHHHHHH Supemegative 57 MSIADLKSRLIKASTSKMTAELTTSKFFNEKDVIRTKIP MLNIAISGAIDGGMQSGLTIFAGPSKSFKSNMSLTMV AAYLNKYPDAICLFYDSEFGITPAYLKSMGVDPERVI HTPIQSVEQLKIDMVNQLETIERGEKVIVFIDSIGNMAS KKETEDALNEKSVADMTRAKSLKSLFRIVTPYFSIKNI PCVAVNHTIETIEMFSKTVMTGGTGVMYSADTVFIIG KRQIKDGSDLQGYQFVLNVEKSRTVKEKSKFFIDVKF DGGIDPYSGLLDMALELGFVVKPKNGWYAREFLDEE TGEMIREEKSWRAKDTNCTTFWGPLFKHQPFRDAIKR AYQLGAIDSNEIVEAEVDELINSEVEEFDSPESDSDEA ADLETDLEQLSDMEEFNEIVEAEVDELINSEVEDFDSP ESDSDEAADLETDLEQLSDMEEFNHHHHHHH UvsX His2 58 MSIADLKSRLIKASTSKMTAELTTSKFFNEKDVIRTKIP MLNIAISGAIDGGMQSGLTIFAGPSKSFKSNMSLTMV 2026234101   18 Sep 2026 AAYLNKYPDAICLFYDSEFGITPAYLKSMGVDPERVI HTPIQSVEQLKIDMVNQLETIERGEKVIVFIDSIGNMAS KKETEDALNEKSVADMTRAKSLKSLFRIVTPYFSIKNI PCVAVNHTIETIEMFSKTVMTGGTGVMYSADTVFIIG KRQIKDGSDLQGYQFVLNVEKSRTVKEKSKFFIDVKF DGGIDPYSGLLDMALELGFVVKPKNGWYAREFLDEE TGEMIREEKSWRAKDTNCTTFWGPLFKHQPFRDAIKR AYQLGAIDSNEIVEAEVDELINSKVEKFKSPESKSKSA ADLETDLEQLSDMEEFNEAGHHHHHPHAHHPLSQSS GHHHHHHHHHHQGYGGSG UvsX+PCF+H is2 59 MSIADLKSRLIKASTSKMTAELTTSKFFNEKDVIRTKIP MLNIAISGAIDGGMQSGLTIFAGPSKSFKSNMSLTMV AAYLNKYPDAICLFYDSEFGITPAYLKSMGVDPERVI HTPIQSVEQLKIDMVNQLETIERGEKVIVFIDSIGNMAS KKETEDALNEKSVADMTRAKSLKSLFRIVTPYFSIKNI PCVAVNHTIETIEMFSKTVMTGGTGVMYSADTVFIIG KRQIKDGSDLQGYQFVLNVEKSRTVKEKSKFFIDVKF DGGIDPYSGLLDMALELGFVVKPKNGWYAREFLDEE TGEMIREEKSWRAKDTNCTTFWGPLFKHQPFRDAIKR AYQLGAIDSNEIVEAEVDELINSKVEKFKSPESKSKSA ADLETDLEQLSDMEEFNEQVQMQLRQVFSQDQQVLQ ERMRYHELQQQQQQQYHETKDMVGSYTQNSNSAIPL FGNNSDTTNQQNSAGHHHHHPHAHHPLSQSSGHHHH HHHHHHQGYGGSG 2026234101   18 Sep 2026 Recombinase loadins protein RPA methods, including those of the present invention, may additionally include / use a recombinase loading protein. 5          Any suitable recombinase loading protein may be used in the RPA methods described herein. Any of the one or more IDR-polypeptides of the present invention may be attached / tethered / tagged to any recombinase loading protein. The recombinase loading protein may originate from a prokaryotic, viral or 10 eukaryotic organism. Exemplary recombinase loading proteins include E. coli RecO, E. coli RecR, UvsY, and mutants or fragments thereof, or combinations thereof. Exemplary UvsY proteins include those derived from myoviridae phages, such as T4, T2, T6, Rb69, Aehl, KVP40, Acinetobacter phage 133, Aeromonas phage 65, cyanophage P-SSM2, cyanophage PSSM4, cyanophage S-PM2, Rbl4, Rb32, 15 Aeromonas phage 25, Vibrio phage nt-1, phi-1, Rbl6, Rb43, Phage 31, phage 44RR2.8t, Rb49, phage Rb3, and phage LZ2. Preferred recombinase loading proteins are UvsY, E. coli RecO, E. coli RecR or any functional variant, analoge, homologe or derivative of any of these proteins. A particularly preferred UvsY recombinase loading protein is Escherichia phage STO 20 UvsY. Any combination of any of these proteins may be used. Preferred concentrations of these proteins is between 0.1-24 pM, 6-24 pM, 4-24 pM and 4-12 pM, preferably about 10 pM, more preferably about 8.6 pM. The recombinase loading protein may be present at between about 0.5 to about 2 times the 25 micromolar concentration of the recombinase agent. Exemplary UvsY recombinase loading proteins tagged with amino acid tag sequences comprising intrinsically disordered regions are set out in Table 22 below. 2026234101   18 Sep 2026 Table 22 Name of protein SEQID NO. Amino acid sequence UvsY [Phage STO] 60 MHHHHHHHMKLEDLQEELDADLAIDMSKLQYETAN NVKLYSKWLRKHSFIRKEMLRIETQKKTALKARLDY YSGRGDGDEFSMDRYEKSEMKTVLAADKDVLKIETT LQYWGILLEFCSGALDAVKSRSFALKHIQDMREFEAG Q UvsY C-Pol CTD 61 MHHHHHHHHKLEDLQEELDADLAIDMSKLQYETAN NVKLYSKWLRKHSFIRKEMLRIETQKKTALKARLDY YSGRGDGDEFSMDRYEKSEMKTVLAADKDVLKIETT LQYWGILLEFCSGALDAVKSRSFALKHIQDMREFEAG QSGSGSGPTSPSYSPTSPSYSPYSPAYS UvsY fib [short] 62 MHHHHHHHHKLEDLQEELDADLAIDMSKLQYETAN NVKLYSKWLRKHSFIRKEMLRIETQKKTALKARLDY YSGRGDGDEFSMDRYEKSEMKTVLAADKDVLKIETT LQYWGILLEFCSGALDAVKSRSFALKHIQDMREFEAG QSGSGSGRGGGFGGRGGFGDRGGRGGRGGFGG UvsY Supl 63 MHHHHHHHHKLEDLQEELDADLAIDMSKLQYETAN NVKLYSKWLRKHSFIRKEMLRIETQKKTALKARLDY YSGRGDGDEFSMDRYEKSEMKTVLAADKDVLKIETT LQYWGILLEFCSGALDAVKSRSFALKHIQDMREFEAG QSGSGYNPQGGYQQNNL UvsY Supl HIS2 64 MAGHHHHHPHAHHPLSQSSGHHHHHHHHHHQGYG GSGKLEDLQEELDADLAIDMSKLQYETANNVKLYSK 2026234101   18 Sep 2026 WLRKHSFIRKEMLRIETQKKTALKARLDYYSGRGDG DEFSMDRYEKSEMKTVLAADKDVLKIETTLQYWGIL LEFCSGALDAVKSRSFALKHIQDMREFEAGQSGSGYN PQGGYQQNNLQ Single strand stabilizing agent RPA methods, including those of the present invention, use a single strand 5 stabilizing agent. Any suitable single strand stabilizing agent (single stranded DNA binding protein) may be used in the RPA methods described herein. Any of the one or more IDR-polypeptides thereof of the present invention may be attached / tethered / tagged to any single strand stabilizing agent. 10           A single strand stabilizing agent is used to stabilize nucleic acids during the various exchange reactions that occur during the RPA reaction. In particular a single strand stabilizing agent is used to stabilize recombinase / ssDNA nucleoprotein filaments. A single strand stabilizing agent can be derived or obtained from any species, e.g., from a prokaryotic, viral or eukaryotic species. 15          Single strand stabilizing agents include single stranded DNA binding proteins from E. coli and those derived from myoviridae phages, such as T4, T2, T6, Rb69, Aehl, KVP40, Acinetobacter phage 133, Aeromonas phage 65, cyanophage P-SSM2, cyanophage PSSM4, cyanophage S-PM2, Rbl4, Rb32, Aeromonas phage 25, Vibrio phage nt-1, phi-1, Rbl6, Rb43, Phage 31, phage 44RR2.81, Rb49, phage Rb3, and 20 phage LZ2. Additional examples of single strand stabilizing agents include A. denitrificans Alide_2047, Burkholderia thailandensis BthaB_33951, Prevotellapollens HMPREF9144_0124, and eukaryotic single stranded DNA binding protein replication protein A. Preferred single strand stabilizing agents are selected from the group consisting 25 of Gp32, E. coli SSB protein, phage T4 Gp32 protein, phage Rb69 Gp32, phage vB_EcoM_NBGl Gp32, and derivatives thereof and any combination thereof. 2026234101   18 Sep 2026 Particularly preferred single strand stabilizing agents are Gp32 and in particular phage vB_EcoM_NBGl Gp32. Any combination of any of these proteins may be used. One preferred concentration of the single strand stabilizing agent is between 5 approximately 5-30 pM, such as approximately 8.6 pM, preferably between approximately 15-25 pM, more preferably approximately 20 pM. Exemplary Gp32 single strand stabilizing agents tagged with amino acid tag sequences comprising intrinsically disordered regions are set out in Table 23 below. 10 Table 23 Name of protein SEQID NO. Amino acid sequence Gp32 (7His) from phage vB 65 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGD YD SCPVCQ YISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADKVASDLDDFDKDMEAFSSAKTEDDFMS SSSSDDSDLDDLLAGLHHHHHHH Gp32Super +ve 66 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGD YD SCPVCQ YISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAAS VAKKVASKLKFKKKMEAFS SAKTKKKFMS S 2026234101   18 Sep 2026 SSSKKSKLKKLLAGLMEAFSSAKTKKKFMSSSSSKK SKLKKLLAGLHHHHHHH Gp32Super -ve 67 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGD YD SCPVCQ YISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADDVASDLDDFDDDMEAFSSADTEDDFMS S S SSDDSDLDDLLAGLMEAFS S ADTEDDFMSS S S SD DSDLDDLLAGLHHHHHHH Gp32- TripleXtail 68 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGD YD SCPVCQ YISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADDVASEFEIVEAEVDELINSKVEKFKSPES KSKSAADLETDLEQLSDMEEFNEIVEAEVDELINSKV EKFKSPESKSKSAADLETDLEQLSDMEEFNEIVEAEV DELINSKVEKFKSPESKSKSAADLETDLEQLSDMEEF NHHHHHH Gp32-fib 69 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGD YD SCPVCQ YISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK 2026234101   18 Sep 2026 KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADKVASDLDDFDKDMEAFSSAKTEDDFMS SSSSDDSDLDDLLAGLPGFSPRGGGFGGRGGFGDRG GRGGRGGFGGGRGRGGGFRGRGRHHHHHHH Gp32-PCF11 70 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGD YD SCPVCQ YISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADKVASDLDDFDKDMEAFSSAKTEDDFMS SSSSDDSDLDDLLAGLNEQVQMQLRQVFSQDQQVL QERMRYHELQQQQQQQYHETKDMVGSYTQNSNSA IPLFGNNSDTTNQQNSHHHHHHH Gp32-Sup 71 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGD YD SCPVCQ YISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADKVASDLDDFDKDMEAFSSAKTEDDFMS SSSSDDSDLDDLLAGLMSDSNQGNNQQNYQQYSQN GNQQQGNNRYQGYQAYNAQAQPAGGYYQNYQGY SGYQQGGYQQYQYNPQGGYQQYNPQGGYQQYNPQ 2026234101   18 Sep 2026 GGYQQQFNPQGGRGNYKNFNYNNNLQGYQHHHHH HH Gp32-Supl 72 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETCSSTHGDYDSCPVCQ YISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADKVASDLDDFDKDMEAFSSAKTEDDFMS SSSSDDSDLDDLLAGLYNPQGGYQQHHHHHHH Gp32-Sup2 73 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGD YD SCPVCQ YISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADKVASDLDDFDKDMEAFSSAKTEDDFMS SSSSDDSDLDDLLAGLYNPQGGYQQYNPQGGYQQH HHHHHH Gp32-Sup3 74 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGD YD SCPVCQ YISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA 2026234101   18 Sep 2026 AAAASVADKVASDLDDFDKDMEAFSSAKTEDDFMS SSSSDDSDLDDLLAGLYNPQGGYQQYNPQGGYQQY NPQGGYQQHHHHHHH Gp32-Sup4 75 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGDYD SCPVCQ YISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADKVASDLDDFDKDMEAFSSAKTEDDFMS SSSSDDSDLDDLLAGLYNPQGGYQQYNPQGGYQQY NPQGGYQQYNPQGGYQQHHHHHHH Gp32-DDX 76 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGDYD SCPVCQ YISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADKVASDLDDFDKDMEAFSSAKTEDDFMS SSSSDDSDLDDLLAGLHHHHHHHFEFMGDEDWEAE INPHMSSYVPIFEKDRYSGENGDNFNRTPASSSEMDD GPSRRDHFMKSGFASGRNFGNRDAGECNKRDNTST MGGFGVGKSFGNRGFSNSRFEDGDSSGFWRESSND CEDNPTRNRGFSKRGGYRDGNNSEASGPYRRGGRG SFRGCRGGFGLGSPNNDLDPDECMQRTGGLFGSRRP VLSGTGNGDTSQSRSGSGSERGGYKGLNEEVITGSG KNS WKSEAEGGES SDTQLEHHHHHH 2026234101   18 Sep 2026 Gp32 PolCTD 77 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGD YD SCPVCQ YISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADKVASDLDDFDKDMEAFSSAKTEDDFMS SSSSDDSDLDDLLAGLHHHHHHHFEFSPFGAYGEAP TSPGFGVSSPGFSPTSPTYSPTSPAYSPTSPSYSPTSPS YSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTS PSYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSP TSPSYSPTSPAYSPTSPSYSPTSPSYSPTSPSYSPTSPSY SPTSPNYSPTSPSYSPTSPGYSPGSPAYSPKQDEQLEH HHHHH Gp32 HNRNPA1 78 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGD YD SCPVCQ YISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADKVASDLDDFDKDMEAFSSAKTEDDFMS SSSSDDSDLDDLLAGLASASSSQRGRSGSGNFGGGR GGGFGGNDNFGRGGNFSGRGGFGGSRGGGGYGGS GDGYNGFGNDGSNFGGGGSYNDFGNYNNQSSNFGP MKGGNFGGRSSGPYGGGGQYFAKPQNQGGYGVSSS S S S YGSGRRFHHHHHHH 2026234101   18 Sep 2026 Gp32 HRP1 79 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGD YD SCPVCQ YISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADKVASDLDDFDKDMEAFSSAKTEDDFMS SSSSDDSDLDDLLAGLGGNNGGNNMNRRGGNFGNQ GDFNQMYQNPMMGGYNPMMNPQAMTDYYQKMQ EYYQQMQHHHHHHH Gp32 HRP2 80 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGD YD SCPVCQ YISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADKVASDLDDFDKDMEAFSSAKTEDDFMS SSSSDDSDLDDLLAGLGGNNGGNNMNRRGGNFGNQ GDFNQMYQNPMMGGYNPMMNPQAMTDYYQKMQ EYYQQMQGGNNGGNNMNRRGGNFGNQGDFNQMY QNPMMGGYNPMMNPQAMTDYYQKMQEYYQQMQ HHHHHHH Gp32 HIS1 81 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGD YD SCPVCQ YISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK 2026234101   18 Sep 2026 VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADKVASDLDDFDKDMEAFSSAKTEDDFMS SSSSDDSDLDDLLAGLPQQNALHHHHGNSSHHHHH HHHHHHHHGQQA GP32 HIS2 82 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGD YD SCPVCQYISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADKVASDLDDFDKDMEAFSSAKTEDDFMS SSSSDDSDLDDLLAGLAGHHHHHPHAHHPLSQSSGH HHHHHHHHHQGYGGSG GP32 HIS3 83 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGD YD SCPVCQYISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADKVASDLDDFDKDMEAFSSAKTEDDFMS SSSSDDSDLDDLLAGLAGHHHHHHHHLPHLPPPHLH HHHHPQHHLHPGSAAAVHPVQQH GP32 HIS4 84 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGD YD SCPVCQ YISKNDLYNTNKTEY 2026234101   18 Sep 2026 SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADKVASDLDDFDKDMEAFSSAKTEDDFMS SSSSDDSDLDDLLAGLNHHASHGHHNSHHPQHHHH HHHHHHHPPPPAPQPPPPPQQQQ GP32 HIS5 85 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGD YD SCPVCQ YISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADKVASDLDDFDKDMEAFSSAKTEDDFMS SSSSDDSDLDDLLAGLSGHHGAHHGAHHPAAAAAY EAFRGPGFAGGGGADDMGAGHHHGAHHAAHHHH AAHHHHHHHHHHGGAGHGGGAGHH Gp32 Mimic 1 86 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGD YD SCPVCQ YISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADKVASDLDDFDKDMEAFSSAKTEDDFMS SSSSDDSDLDDLLAGLYDPTSPSYDPTSPSYDPTSPSY DPTSPSYDPTSPSYDPTSPSYDPTSPSHHHHHHH 2026234101   18 Sep 2026 Gp32 Mimic2 87 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGD YD SCPVCQ YISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADKVASDLDDFDKDMEAFSSAKTEDDFMS SSSSDDSDLDDLLAGLYSPTDPSYSPTDPSYSPTDPSY SPTDPSYSPTDPSYSPTDPSYSPTDPSHHHHHHH Gp32 short pol ctd 88 MFKRKSTADLAAQMAKLNGNKGFSSEDKGEWKLK LDASGNGQAVIRFLPAKTDDALPFTILVNHGFKKNG KWYIETC S STHGD YD SCPVCQ YISKNDLYNTNKTEY SQLKRKTSYWANILVVKDPQAPDNEGKVFKYRFGK KIWDKINAMIAVDTEMGETPVDVTCPWEGANFVLK VKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQM VDLSEMTSKDKFKSFEELNTKFNQVLGTAALGGAA AAAASVADKVASDLDDFDKDMEAFSSAKTEDDFMS SSSSDDSDLDDLLAGLTSPSYSPTSPSYSPTSPGYSPT SPAYSPTSPTYSPTSPTYSPHHHHHHH Polymerase RPA methods, including those of the present invention, use a polymerase. 5          Any suitable polymerase may be used in the methods described herein. Any of the one or more IDR-polypeptides of the present invention may be attached / tethered / tagged to any suitable polymerase. 2026234101   18 Sep 2026 For the synthesis or amplification of DNA, DNA polymerase are preferably used. One advantage of the RPA reaction is that there is no limit on the type of polymerase that can be used. For example, eukaryotic, prokaryotic and bacteriophage 5 polymerases can be used. The DNA polymerase may be a eukaryotic polymerase. Examples of eukaryotic polymerases that may be used include pol-a, pol-0, pol-8, pol-s or any functional variant, analoge, homologe or derivative thereof and any combination thereof. The DNA polymerase may be a prokaryotic polymerase. Examples of 10   prokaryotic polymerases that may be used include E. coli DNA, polymerase I Klenow fragment, E. coli DNA polymerase I, E. coli DNA polymerase II, E. coli DNA polymerase III, E. coli DNA polymerase IV, E. coli DNA polymerase V, Bacillus stearothennophilus polymerase I large fragment, Bacillus subtilis Pol I large fragment (Bsu polymerase), Listeria monocytogenes DNA polymerase I, Staphylococcus aureus 15 DNA polymerase 1 (Sau) or any functional variant, analoge, homologe or derivative thereof and any combination thereof. The DNA polymerase may be a bacteriophage polymerase. Examples of bacteriophage polymerases that may be used in the methods described herein include Phi-29 DNA polymerase, T7 DNA polymerase, bacteriophage T4 gp43 DNA 20 polymerase, or any functional variant, analoge, homologe or derivative thereof and any combination thereof. The DNA polymerase typically contains strand displacing properties. DNA polymerases can use the free 3’-hydroxyl of the invading strand to catalyze DNA synthesis by incorporation of new nucleotides. A number of polymerases 25 can use the 3 ’-hydroxyl of the invading strand to catalyze synthesis and simultaneously displace the other strand as synthesis occurs. For example E. coli polymerase II or III can be used to extend invaded D-loops. In addition, E. coli polymerase V normally used in SOS-lesion-targeted mutations in E. coli can be used. All of these polymerases can be rendered highly processive through their interactions and co-operation with the 30 (3-dimer clamp, as well as single stranded DNA binding protein (SSB) and other 2026234101   18 Sep 2026 components. Other polymerases from prokaryotes, viruses, and eukaryotes can also be used to extend the invading strand. Many DNA polymerases possess 3’-5’ exonuclease activity, and some also possess 5’-3’ exonuclease activity, which is undesirable in RPA reactions as it results in 5 digestion of one DNA strand progressively as the polymerase moves forward, rather than displacement. The 3’-5’ exonuclease has potential advantages as well as its obvious disadvantages. On the one hand 3’-5’ exonuclease activity increases the fidelity of the replication reaction, and can also prevent stalling of polymerases at points of 10 misincorporation. High fidelity amplification is desirable for many DNA applications. The 3’-5’ exonuclease activity may also be appropriate for amplification of larger DNA fragments where stalling due to misincorporation could inhibit effective amplification. Despite these clear advantages of 3’-5’ exonuclease activity there are some disadvantages. The free oligonucleotides can be subject to end-dependant degradation 15 when polymerases possessing 3’-5’ exonuclease are employed. Reaction noise can be reduced by utilising polymerases lacking 3’-5’ exonuclease activity. This suggests mispriming may result from oligonucleotides that have been shortened by the 3’-5’ exonuclease activity of polymerases. Consequently 3’-5’ exonuclease editing activity, pyrophosphorylysis, or any other similar editing 20 activity can be a source of noise. This can be suppressed to a large extent by using saturating amounts of relatively cooperative Gp32 protein with some polymerases such as the Klenow fragment. Nevertheless, polymerases for use in the methods described herein might be provided which lack 3’-5’ exonuclease activity. The DNA polymerase may be present at a concentration of between 10,000 25 units / ml to 10 units / ml, such as between 5000 units / ml to 500 units / ml. Accessory agents RPA reactions, including those of the present invention, may further utilize 30 accessory agents. 2026234101   18 Sep 2026 Any of the one or more IDR-polypeptides of the present invention may be attached / tethered / tagged to any accessory agent. These accessory agents include a single strand binding protein, a helicase, a topoisomerase, a resolvase and any combination thereof. Such agents may possess 5 unwinding, relaxing, and resolving activities respectively on nucleic acids. The accessory agents may also include RuvA, RuvB, RuvC, RecG, PriA, PriB, PriC, DnaT, DnaB, DnaC, DnaG, DnaX clamp loader, polymerase core complex, DNA ligase and a sliding clamp and any combination thereof. The sliding clamp may be E. coll p-dimer sliding clamp, the eukaryotic PCNA sliding clamp, or the T4 sliding clamp 10   gp45 and a combination thereof. The accessory agents may include, in addition, DNA Polymerase III holoenzyme complex consisting of P-Clamp, DnaX Clamp Loader, and the Polymerase Core Complex. These latter accessory agents would allow the performance of leading and lagging RPA. RPA reactions may be performed with one or more additional enzymes that can 15 promote efficient disassembly of recombinase agent / dsDNA complexes after initiation of DNA synthesis. These enzymes include those that are capable of stimulating 3’ to 5’ disassembly and those capable of supporting 5’ to 3’ disassembly. Such additional enzymes include several polymerases that can displace RecA in the 3’ to 5’ direction and can stimulate 3’ to 5’ disassembly of recombinase 20 agent / dsDNA complexes. These DNA polymerases include E. coll PolV and homologous polymerase of other species. Inclusion of E. coli PolV or any functional variant, analoge, homologe or derivative thereof may improve the amplification efficiency. Other enzymes include a class of enzymes called helicases that can be used to 25 promote the disassembly of RecA from dsDNA. These promote disassembly in both the 5’ to 3’ and 3’ to 5’ directions. An ideal helicase complex for stimulating disassembly of RecA from intermediates consists of the E. coli proteins RuvA and RuvB. The RuvAB complex promotes branch migration, and dissociates the RecA protein, allowing RecA to be recycled. Incorporation of RuvAB into the RPA mixture 30 can promote the dissociation of RecA from dsDNA following strand exchange and displacement, allowing renewed synthesis of the duplicated template from the same site. 2026234101   18 Sep 2026 Additionally, the RuvAB complex can act in concert with RuvC, which finally cuts and resolves Holliday junctions. With RuvC added to the RPA reaction mixture, complicated structures such as Holliday junctions formed at invasion sites, can be resolved. 5          Still other enzymes include the E. coli RecG protein. RecG can stimulate disassembly of branch structures. Other enzymes useful in an RPA reaction mixture are those that allow continual generation of RecA nucleoprotein filaments in the presence of ATP and the single strand stabilizing agent. Accordingly, RecO and RecR, and optionally RecF proteins 10 may be used. Exonuclease enzymes are often included in RPA reaction mixtures. These are included for the efficient operation of cleavable probes. One example of an exonuclease enzyme commonly used is Exonuclease III. Any of the IDR polypeptides of the present invention may be attached / tethered / tagged to any exonuclease. 15 Primers RPA methods employ polymerases to generate copies of template nucleic acid molecules. RPA methods, including those of the present invention, therefore use 20 primers to initiate extension by polymerases. It is a necessity of most nucleic acid polymerases that incorporation requires a free 3’-hydroxyl moiety on the terminal sugar of a short stretch of double stranded nucleic acid adjacent to the site of new synthesis. This stretch of double stranded nucleic acid is typically formed on a template by a short oligonucleotide typically 25 having a complementary sequence, called a primer, which serves as an initiation site for the polymerase synthesis reaction. In some cases a 3 ’ modification, such as a sulfydryl, may be utilized to prime the synthesis reaction. The primer nucleic acid, which is basepaired with the template and extended by the polymerase, can be RNA or DNA. Typically, for in vitro reactions the primer is supplied as a short, often chemically 30 synthesized, single stranded DNA (or modified DNA or RNA), and is usually referred to as an oligonucleotide primer. The primer is often of a specific sequence, although 2026234101   18 Sep 2026 random primers can also be used. The primer is targeted to complementary sequences by virtue of its specific base-pairing capacity. Formation of hybrids between the oligonucleotide primer and target nucleic acid are typically formed by incubation of the two in solution under conditions of salt, pH, and temperature that allow spontaneous 5 annealing. The primers used in RPA may have a single stranded region for hybridization to the target DNA in the presence of a recombinase agent. The single stranded region may be, for example, about 10 bases, about 15 bases, about 20 bases, about 25 bases, about 30 bases, about 40 bases, and about 50 bases. Even longer regions such as about 75 10 bases, about 100 bases, about 150 bases or more may in theory be used. The choice of single stranded regions will depend on the complexity of the starting nucleic acid so that for example, a human genome may require a longer primer while a plasmid may require a much shorter primer. A preferred primer length is between about 30 to about 50 bases. For example, 15 between 30 to 45 bases, between 30 to 40 bases, between 30 to 35 bases, between 35 to 40 bases, between 40 to 45 bases, and between 45 to 50 bases. While the abovereferenced primer lengths are indicated, a recombinase and / or single stranded binding protein with an optimum primer length of less than 30 bases is also possible and envisioned. 20         The primers used in RPA are preferably DNA although PNA, and RNA are also suitable for use as primers. It is noted that in fact, in natural DNA replication, DNA polymerases elongate genomic DNA by extension from RNA primers. Primers may be synthesized according to standard techniques. Modified bases and / or linker backbone chemistries may be desirable and functional in some cases. 25 Additionally oligonucleotides maybe modified at their ends, either 5’ or 3’, with groups that serve various purposes e.g. fluorescent groups, quenchers, protecting (blocking) groups (reversible or not), magnetic tags, proteins etc. In some cases single stranded oligonucleotides may be used for strand invasion, in others only partly single stranded nucleic acids may be used, the 5’ stretch of sequence of an invading nucleic acid being 30 already hybridized to an oligonucleotide. 2026234101   18 Sep 2026 The primers may comprise a 5 ’ region that is not homologous to the target nucleic acid. It should be noted that amplification may be achieved even if the primers are not completely complementary to the target nucleic acid. The primers may be non-complementary by having additional sequences at their 5’ ends. These additional 5 sequences may be, for example, the sequence for a restriction endonuclease recognition site or the sequence that is complementary to a sequencing primer. The restriction endonuclease recognition site may be useful for subsequent cleavage of the amplified sequence. The use of restriction endonuclease that cleaves nucleic acid outside the restriction endonuclease recognition site is also contemplated. The sequence that is 10   complementary for a sequencing primer may allow rapid DNA sequencing of the amplified product using commercially available primers or commercially available sequencing apparatus. Software to design oligonucleotides for use in in vitro DNA synthesis reactions is well established, particularly for use in PCR. The considerations for the RPA method 15   are similar and include the optimisation of the melting temperature of the oligonucleotide, avoidance of hairpin formation within an oligonucleotide and selection against complementarity with other oligonucleotides present in a given reaction. It is therefore important to design oligonucleotide primer pairs to avoid undesirable side reactions. 20          Besides optimising oligonucleotide sequence design there are additional approaches to reduce or eliminate primer dimer formation. As noted elsewhere herein, reaction noise can be reduced by utilising polymerases lacking 3’-5’ exonuclease activity. This suggests mispriming may result from oligonucleotides that have been shortened by the 3’-5’ exonuclease activity of polymerases. Consequently 3’-5’ 25 exonuclease editing activity, pyrophosphorylysis, or any other similar editing activity can be a source of noise. In addition to using polymerases lacking exonuclease activity and the removal of pyrophosphate with pyrophosphatase, use of synthetic oligonucleotides with a non-hydrolysable backbone at the ultimate and / or penultimate link may be beneficial to reduce reaction noise. Alternative backbones could be 30 selected from the considerable range of chemistries available such as phosphorothiorate, morpholino, locked nucleic acid, or peptide nucleic acid. 2026234101   18 Sep 2026 Reagents for use in RPA reactions Reagents for use in RPA methods, including those of the present invention, are 5 outlined below. dNTPs dNTPs, for example dATP, dGTP, dCTP, and dTTP, and derivatives and 10 analogs thereof, may be added to an RPA reaction. In leading and lagging strand RPA, ATP, GTP, CTP, and UTP may also be included for synthesis of RNA primers. In addition, ddNTPs (ddATP, ddTTP, ddGTP and ddGTP and derivatives and analogs thereof) may be used to generate fragment ladders. The dNTP maybe used at a concentration of between 1 mM to 200 mM of each 15 NTP species. A mixture of dNTP and ddNTP may be used with ddNTP concentrations at 1 / 100 to 1 / 1000 of that of the dNTP (1 mM to 200 mM). The RPA may be performed in the presence of ATP, a hydrolysable ATP analog, or another nucleoside triphosphate. The ATP analog may be, for example, 20 dATP, ddATP, or another nucleoside triphosphate analog such as UTP. Reducing agents Reducing agents which may be used in the RPA reaction include DTT. The 25 DTT concentration may be between 1 mM and 10 mM, preferably 1 mM. ATP ATP or an ATP analog may be used in the RPA reaction. 2026234101   18 Sep 2026 The ATP or ATP analog may be any of ATP, ATP-y-S, ATP-^-S, ddATP or a combination thereof. A preferred ATP or ATP analog concentration is between 1 mM and 10 mM, preferably 2.5 mM. 5   System for A TP regeneration Other components of the RPA reaction may include a system for ATP regeneration (i.e. a system to convert ADP to ATP). Such a system may be, for example, phosphocreatine and creatine kinase. 10          An ATP regeneration system permits persistent recombination reactions, as recombinases have an extremely high rate of ATP hydrolysis when bound to nucleic acids. In particular, the UvsX protein has a hydrolysis rate 10-20 times higher than RecA and can consume 200 molecules of ATP per minute per monomer. A number of systems are available. The creatine kinase / phosphocreatine system is preferred. When 15 UvsX is employed the AMP that is produced may be converted into ATP. Chicken myokinase may additionally be used, which converts a molecule of AMP and one of ATP to two molecules of ADP. ADP can then be converted to ATP using the creatine kinase / phosphocreatine system. Poor regeneration of ATP can reduce the reaction rate. In the RPA methods described herein phosphocreatine is preferably used at a 20 concentration of between 15-25 mM, more preferably 20 mM. Creatine kinase is preferably used at a concentration of between about 0.25-5.0 pM, more preferably 1 pM. Multivalent metal cations 25 The buffer solution in an RPA reaction preferably contains multivalent metal cations. The buffer may contain a functional equivalent of a multivalent metal cation. The buffer solution in an RPA reaction more preferably contains divalent metal cations. The buffer may contain a functional equivalent of a divalent metal cation. 30          Any suitable multivalent or divalent metal cation or functional equivalent thereof may be used, either as a single agent or combination of agents. 2026234101   18 Sep 2026 The specific multivalent or divalent metal cation or functional equivalent thereof which achieves optimal results in promoting / enhancing IDR-mediated phase separation in an RPA reaction, as well as the specific concentration of the multivalent / divalent metal cation used may depend upon the specific IDR polypeptide which is used. The 5 optimal multivalent / divalent metal cation or functional equivalent thereof, and the optimal concentration thereof, can be established empirically using routine tests, including RPA reactions themselves and / or the phase separation assays which are described further herein. The divalent metal cation may be Mg2+, Mn2+, Ca2+, Co2+, Ni2+ or Cu2+. Any of 10   these cations may be used as a single agent, or any combination of cations may be used. Preferably they are used as single agents. Preferred divalent metal cations are Mg2+, Mn2+ and Ca2+. A particularly preferred divalent metal cation is Mg2+. A preferred concentration range is from 30 to 40 mM, more preferably from 33 to 39 mM. 15          The buffer may contain Mg2+ ions, preferably at the indicated concentrations. More preferably the buffer contains MgOAc at the indicated concentrations. The buffer may contain Ca2+ ions, preferably at the indicated concentrations. More preferably the buffer contains CaCh at the indicated concentrations. The buffer may contain Mn2+ ions, preferably at the indicated concentrations. 20 More preferably the buffer contains MnCh at the indicated concentrations. Buffers The buffer solution in an RPA reaction may be a Tris-HCl buffer, a Tris-Acetate 25 buffer, or a combination thereof. The buffers may be present at a concentration of between about 10 mM to about 100 mM. A preferred buffer is a Tris-HCl buffer used at a concentration of between about 20 mM to about 30 mM, most preferably 25 mM. The buffered pH may be between 6.5 to 9.0, preferably pH 8.3. The buffer may contain potassium acetate between about 5 mM to about 50 mM, 30 preferably between about 10 mM to about 40 mM. 2026234101   18 Sep 2026 Reaction components A preferred but non-limiting set of reaction components for an RPA reaction is as follows below. 5 Tris HC1 pH 8.3 KOAc DTT ATP 25 mM 7.5 mM 1 mM 2.5 mM 10 Phosphocreatine 20 mM Creatine kinase 1 pM dNTPs 1 mM Gp32 20 pM UvsX 4.8 pM 15 UvsY 8.6 pM 5*. aureus DNA polymerase 1 (Sau) Or B. subtilis DNA polymerase 1 (Bsu) 0.135 pM Exonuclease III 0.27 pM MgOAc 33 mM 20 Forward primer 0.4 pM Reverse primer 0.4 pM Exo probe 0.12 pM RPA reaction conditions RPA reactions, including those of the present invention, may incubated for any suitable length of time. Any of the RPA reactions may be incubated for between 5 minutes and 16 hours or more, such as between 15 minutes and 3 hours or between 30 minutes and 2 hours. 2026234101   18 Sep 2026 The incubation may be performed until a desired degree of amplification is achieved. The desired degree of amplification may be 10 fold, 100 fold, 1000 fold, 10,000 fold, 100,000 fold or 1,000,000 fold amplification. One benefit of RPA is that the reaction may be performed at reduced 5 temperatures compared to techniques which require thermal cycling, such as PCR. A further advantage of RPA is that the temperature is not critical and precise control, while preferred, is not absolutely necessary. For example, in a field environment, it is sufficient to incubate the RPA reaction at room temperature, or close to body temperature (35°C to 38°C), for example by placing the sample in a body crevice. 10   Furthermore, the RPA reaction may be performed without temperature induced melting of the template nucleic acid. Thus any of the RPA reactions may be performed at any suitable temperature. The RPA reactions may be performed at less than 45°C. The RPA reactions may be performed at less than 40°C. The RPA reactions may be performed at less than 15   35°C. The RPA reactions may be performed at less than 30°C. The RPA reactions may be performed at between 20°C and 50°C, between 20°C and 40°C, such as between 20°C and 30°C. Freeze drying of RPA reaction components 20 One advantage of RPA reactions is that the reagents, with the possible exception of the crowding agent (if used) and buffer, may be freeze dried (i.e., lyophilized) before use. Freeze dried reagents offer the advantage of not requiring refrigeration to maintain activity. For example, a tube of RPA reagents may be stored at room temperature. This 25 advantage is especially useful in field conditions where access to refrigeration is limited. RPA reagents may be freeze dried onto the bottom of a tube, or on a bead or any other suitable type of solid support. To perform an RPA reaction the freeze dried reagents are reconstituted in a buffer solution and with a crowding agent (if used), or 30 simply a buffered solution or water, depending on the composition of the freeze-dried reagents. Then a target nucleic acid, or a sample suspected to contain a target nucleic 2026234101   18 Sep 2026 acid is added. The reconstitution liquid may also contain the sample nucleic acid. The reconstituted reaction is incubated for a period of time and the amplified nucleic acid, if present, is detected. In any one of the RPA methods described herein, the reagents that can be freeze 5 dried before use include, at least, the recombinase agent, the recombinase loading protein, the single strand stabilizing agent, the DNA polymerase, the dNTPs or the mixture of dNTPs and ddNTPs, the reducing agent, the ATP or ATP analog, primers and probe. Stabilizing agents such as trehalose sugar may be included in the freeze dried 10 mixture, for example at 20 mM to 200 mM and most optimally 40 mM to 80 mM in the reconstituted reaction, in order to improve freeze-drying performance and shelf life. If desired, the freeze dried reagents may be stored for 1 day, 1 week, 1 month or 1 year or more before use. Biochemical reaction reagents, such as RPA reagents, may be freeze dried 15   together with a crowding agent. However, complex inter-related issues may exist which may justify the omission of a crowding agent in a lyophilized mixture. For example, the user may experience difficulty in the effective rehydration of the freeze dried crowding agent, or the user may experience other detrimental effects, including the need for larger lyophilized pellets. Accordingly, there may be advantages in being able to exclude 20 some or all of a crowding agent from freeze dried materials which include among other things reduction of pellet size, shorter cycle times, and easier rehydration. However this has the consequential disadvantage that a crowding agent, if used, would need to be added fresh prior to use after the biochemical reaction mixtrure was rehydrated and prepared for use. This could be problematic in certain situations, such as for point-of- 25 care use or field use. An advantage of the IDR-based reagents of the present invention is that they would not be expected to exhibit the same drawbacks as crowding agents in a lyophilized setting, and could therefore readily be freeze dried with other biochemical reaction components, thus obviating the need to add fresh additional reagents prior to use. 30 2026234101   18 Sep 2026 Detection ofRPA reaction products Detection of RPA reaction products may be performed using any suitable method. 5          For example, detection may be performed using electrophoresis on an agarose or PAGE gel followed by ethidium bromide staining. Monitoring a RPA reaction may involve, for example, removing a fraction of an RPA, reaction, isolating the unincorporated fraction, and detecting the unincorporated primer. Since the size of an unincorporated primer may be less than 50 bp, less than 40 10 bp, less than 30 bp or less than 25 bp, and the size of the amplified product may be greater than 1Kb, greater than 2 Kb, greater than 5 Kb, or greater than 10 Kb, there is a great size difference between the incorporated and unincorporated primer. The isolation of the unincorporated primer may be performed rapidly using size exclusion chromatography such as, for example, a spin column. If a primer is labeled, a monitor 15 procedure comprising a spin column and a measurement (e.g., fluorescence or radioactivity) can be performed in less than one minute. Another alternative for separating elongated primers from unelongated primers involve the use of PAGE. For example, the elongated primer may be separated from the unelongated primer by gel electrophoresis in less than 5 minutes. 20          Yet another alternative for separating elongated primers involves the use of immobilized oligonucleotides. For example oligonucleotides homologous to sequences found uniquely within the amplified DNA sequence can be used to capture nucleic acids produced by primer elongation specifically. These capturing oligonucleotides can be immobilized on a chip, or other substrate. Capture of the elongated oligonucleotides by 25 the capturing oligonucleotides can be performed by RecA protein mediated methods, or by traditional solution hybridizations if necessary. The use of fluorescent probes is most commonly used and preferred for the detection of RPA amplification products and have the advantage of providing for realtime detection. 30          These probes are labelled with a fluorophore, such as fluorescein (FAM) and a quencher, such as a Black Hole Quencher, in close proximity to the fluorophore. The 2026234101   18 Sep 2026 probes have a blocking group at the 3 ’ end to prevent extension from the probe by polymerase. A fluorescent signal is detected when the probe is cleaved and when the quencher and fluorophore are separated, allowing for real-time detection. The probe contains an abasic site, typically tetrahydrofuran (THF) or dR group, and cleavage 5 occurs at the abasic site, typically by E. coli Exonuclease III (cleaving at THF) or E. coli fpg (glycolyase / lyase) (cleaving at the dR group). Kits comprisinz RPA reaction components 10          The invention also provides a kit for performing an RPA reaction. The kit may comprise any of the reagents described herein for RPA in any one of the concentrations described above. The kit may comprise any of the IDR-tagged macromolecules and / or IDR-tagged polypeptides described and defined herein. Preferably the kit further comprises 15 additional RPA components selected from an RPA recombinase agent, and / or an RPA recombinase loading protein, and / or polymerase, and / or first and second nucleic acid primers, and / or an exonuclease, and / or a buffer, and / or a source of multivalent metal ions, preferably divalent metal cations such as Mg2+, Mn2+, Ca2+, Co2+ orNi2+,. The reagents of the kit may be freeze dried, in which case the reagents may be 20 provided in any suitable amount such that when reconstituted the appropriate reagent concentration is achieved. Polymerase 25          As discussed above, any of the IDR amino acid sequences as described and defined herein may be tagged to any protein component required for the performance of a nucleic acid synthesis reaction. Any of the IDR amino acid sequences as described and defined herein may be tagged to any protein component required for the performance of a nucleic acid 30 synthesis reaction wherein a polymerase is used to synthesize a new nucleic acid molecule by extending a primer nucleic acid molecule. 2026234101   18 Sep 2026 Accordingly, any suitable polymerase may be tagged with an IDR amino acid sequence as described and defined herein. The polymerase may be one which is compatible with and may be used in any reaction used to synthesize a new nucleic acid molecule by extending a primer nucleic acid molecule. 5         The polymerase may be one which is compatible with and may be used in any nucleic acid amplification reaction. The nucleic acid amplification reaction may be a reaction which involves thermal cycling. The nucleic acid amplification reaction may be an isothermal amplification reaction. The nucleic acid amplification reaction may be polymerase chain reaction (PCR), polymerase spiral reaction (PSR), loop mediated 10 isothermal amplification (LAMP), nucleic acid sequence based amplification (NASBA), self-sustained sequence replication (3SR), rolling circle amplification (RCA), strand displacement amplification (SDA), multiple displacement amplification (MDA), ligase chain reaction (LCR), helicase dependant amplification (HDA), ramification amplification method (RAM), recombinase polymerase amplification (RPA), 15   transcription-mediated amplification (TMA) or nicking enzyme amplification reaction (NEAR). Sequence tags 20         Any of the IDR-macromolecules or any of the IDR-polypeptides involved in biochemical reactions as described herein, including those involved in RPA reactions, may comprise one or more sequence tags. If used, any such sequence tags are preferably attached to a polypeptide as a fusion protein, as described herein. Sequence tags and means for attaching sequence tags to polypeptides are well known in the art. 25          Sequence tags may be short amino acid sequences or larger polypeptides including proteins. Sequence tags may be attached to the C-terminus of a polypeptide, to the N-terminus of a polypeptide tag, to both the C- and N-terminus of a polypeptide or at any amino acid position along the length of a polypeptide in any combination. 30          Non-limiting examples of suitable amino acid sequence tags include 6-histidine (6X-His; HHHHHH; SEQ ID NO:89), c-myc epitope (EQKLISEEDL; SEQ ID NO:90), 2026234101   18 Sep 2026 FLAG® octapeptide (DYKDDDDK; SEQ ID NO:91), Protein C (EDQVDPRLIDGK; SEQ ID NO:92), Tag-100 (EETARFQPGYRS; SEQ ID NO:93), V5 epitope (GKPIPNPLLGLDST; SEQ ID NO:94), VSV-G (YTDIEMNRLGK; SEQ ID NO:95), Xpress (DLYDDDDK; SEQ ID NO:96), and hemagglutinin (YPY-DVPDYA; SEQ ID 5   NO:97). Non-limiting examples of suitable protein tags include P-galactosidase, thioredoxin, His-patch thioredoxin, IgG-binding domain, inteinchitin binding domain, T7 gene 10, glutathione-S-transferase (GST), green fluorescent protein (GFP), and maltose binding protein (MBP). 10          It will be understood by those in the art that sequence tags and protein tags can be used interchangeably, e.g., for purification and / or identification purposes. Solid phase biochemical reactions 15          Biochemical reactions performed in processes according to the invention may be performed using solid phase or reversible solid phase techniques. Solid phase reaction systems suitable for performing the processes, uses and methods described herein may comprise a surface. Any suitable surface may be used. Data described herein demonstrate that biochemical reactions may be performed 20 with IDR-based reagents according to the invention using solid phase techniques in the absence of a crowding agent. One particular example is recombinase polymerase amplification of nucleic acids wherein primers are attached to a solid surface. Any suitable biochemical reaction amenable to performance using solid phase methods may be performed using such methods using processes according to the invention involving 25 any of the IDR-based reagents described and defined herein. A variety of such solid phase techniques are known in the art and may be used. Macromolecules, such as polynucleotides, including nucleic acid amplification primers, peptides, haptens, hormones, drugs, etc. may be immobilized to a surface. Any suitable macromolecular component of a biochemical reaction may be 30 immobilized to a surface, including the IDR-based reagents described and defined herein. 2026234101   18 Sep 2026 Macromolecules, such as polynucleotides, e.g. primers for use in amplification reactions, may be immobilized to surfaces directly or indirectly. For example they may be attached directly to surfaces by chemical bonding. They may be indirectly attached to surfaces via an intermediate surface. 5          A surface may be e.g. a planar surface such as glass, a gel-based material, or the surface of a microparticle such as a bead or functionalised quantum dot. The material comprising the surface may itself be bound to a substrate. The substrate may be comprise any suitable material such as glass, plastics or polymeric material. Macromolecules involved in biochemical reactions according to processes of the 10 invention may be immobilized to a gel-based material such as e.g. polyacrylamide or a hydrogel, and wherein the gel-based material is itself bound to a supporting substrate such as glass or a plastics or polymeric material. Pre-formed polynucleotides, for example, can be immobilized to surfaces by methods commonly employed to create nucleic acid microarrays. For example, 15   polynucleotides may be synthesized and then spotted or printed onto a surface, typically a planar surface. Polynucleotides may be deposited onto surfaces using contact printing techniques. For example, solid or hollow tips or pins may be dipped into solutions comprising pre-formed polynucleotides and contacted with the surface. Alternatively, polynucleotides may be adsorbed onto micro-stamps and then transferred to a surface 20 by physical contact. Non-contact printing techniques include thermic printing or piezoelectric printing wherein sub-nanolitre size microdroplets comprising pre-formed polynucleotides may be ejected from a printing tip using methods similar to those used in inkjet and bubblejet printing. Polynucleotides may be synthesised directly on surfaces such as using so-called 25 “on-chip” methods employed to create nucleic acid microarrays. On-chip techniques for generating polynucleotides include photolithography which involves the use of UV light directed through a photolithographic mask to selectively activate a protected nucleotide allowing for the subsequent incorporation of a new protected nucleotide. Cycles of UV-mediated deprotection and coupling of pre-determined nucleotides allows 30 the in situ generation of a polynucleotide having a desired sequence. As an alternative to the use of a photolithographic mask, polynucleotides may be created on surfaces by 2026234101   18 Sep 2026 the sequential deposition of nucleobases using inkjet printing technology and the use of cycles of coupling, oxidation and deprotection to generate an oligonucleotide having a desired sequence (for a review see Kosuri and Church, Nature Methods, 2014, 11, 499507). 5          Surfaces for the attachment of macromolecules, including polynucleotides, peptides, haptens, hormones, drugs, etc. can be made of any suitable material. Typically a surface may comprise silicon, glass or any suitable polymeric material, such as polystyrene. A surface may comprise a gel surface, such as a polyacrylamide surface or a hydrogel surface. The gel surface may in turn be coupled to or bound to a solid 10 support or a substrate, said support or substrate may comprise any suitable material such as silicon, glass or any suitable polymeric material. A surface may comprise a hydrogel material coupled to a polystyrene material. A surface may be the surface of a microparticle, often referred to as a microsphere or microbead, or simply a bead. 15          A surface may comprise a hydrogel material coupled to a polystyrene material in the form of a microbead. A variety of surface attachment methods and chemistries are available for the immobilization of macromolecules, such as polynucleotides, to surfaces such as microbeads. Surfaces may be functionalised or derivatized to facilitate attachment. 20 Such functionalisations are known in the art. For example, a surface may be functionalised with a polyhistidine-tag (hexa histidine-tag, 6xHis-tag, His6 tag or His-tag®), Ni-NTA, streptavidin, biotin, an oligonucleotide, a polynucleotide (such as DNA, RNA, PNA, GNA, TNA or LNA), carboxyl groups, quaternary amine groups, thiol groups, azide groups, alkyne groups, DIBO, lipid, FLAG-tag (FLAG octapeptide), 25 polynucleotide binding proteins, peptides, proteins, antibodies or antibody fragments. The surface may be functionalised with a molecule or group which specifically binds to the macromolecule to be immobilized or to another moiety attached to the macromolecule to be immobilized. Covalent immobilization of macromolecules to surfaces is commonly employed. Purely by way of example, a carboxylate-modified 30 polystyrene latex surface is suitable for the covalent attachment of e.g. amine-terminated proteins, DNA, or other molecules for example via EDAC-mediated 2026234101   18 Sep 2026 coupling. Other techniques are available. Although macromolecules will typically be attached chemically, they may also be attached to surfaces by indirect means such as via affinity interactions. For example, macromolecules to be immobilized may be functionalised with biotin and bound to surfaces coated with avidin or streptavidin, or 5 vice-versa. In any of the processes, uses and methods described and defined herein, a macromolecule may be attached to a surface via one or more covalent bonds. The one or more covalent bonds may be formed between a functional group on the surface and a functional group on the marcomolecule. The functional group on the macromolecule 10 may be e.g. an amine group, a thiol group, a thiophosphate group or a thioamide group. The functional group on the surface may be e.g. a bromoacetyl group, optionally wherein the bromoacetyl group is provided on a polyacrylamide surface derived using N- (5- bromoacetamidylpentyl) acrylamide (BRAPA). In any of the processes, uses and methods described and defined herein a 15   macromolecule may be attached to a surface, either directly or indirectly, via a linker. Any suitable linker which is biocompatible in nature may be used. A linker may be a linear linker or a branched linker. A linker may comprise a hydrocarbon chain. A hydrocarbon chain may comprise from 2 to about 2000 or more carbon atoms. The hydrocarbon chain may 20 comprise an alkylene group, e.g. C2 to about 2000 or more alkylene groups. The hydrocarbon chain may have a general formula of -(CH2)n- wherein n is from 2 to about 2000 or more. The hydrocarbon chain may be optionally interrupted by one or more ester groups (i.e. -C(O)-O-) or one or more amide groups (i.e. -C(O)-N(H)-). Any linker may be selected from the group comprising polyacrylamide, poly(2-25 hydroxyethyl methacrylate), Poly-2-methyl-2-oxazoline (PMOXA), zwitterionic polymers, e.g. poly(carboxybetaine methacrylate) (PCBMA), poly[ N -(3-sulfopropyl)-N-methacryloxyethyl- N, N dimethyl ammonium betaine] (PSBMA), glycopolymers, and polypeptides. A linker may comprise oligoethylene glycol-phosphate units having a general 30 formula of -[(CH2-CH2-O)n-PO2'-O]m- where n is from 1 to about 600 or more and m could be 1-200 or more. 2026234101   18 Sep 2026 Any of the above-described linkers may be attached at one end of the linker to a macromolecule described herein, and at the other end of the linker to a first functional group wherein the first functional group may provide a covalent attachment to a surface. The first functional group may be e.g. an amine group, a thiol group, a thiophosphate 5 group or a thioamide group as further described herein. The surface may be functionalised with a further functional group to provide a covalent bond with the first functional group. The further functional group may be e.g. a 2-bromoacetamido group as further described herein. Optionally a bromoacetyl group is provided on a polyacrylamide surface derived using N-(5-bromoacetamidylpentyl) acrylamide 10 (BRAPA). The further functional group on the surface may be a bromoacetyl group, optionally wherein the bromoacetyl group is provided on a polyacrylamide surface derived using N-(5-bromoacetamidylpentyl) acrylamide (BRAPA) and the first functional group may be e.g. an amine group, a thiol group, a thiophosphate group or a thioamide group as appropriate. The surface to which polynucleotides are attached may 15 comprise a gel. The surface may comprise a polyacrylamide surface, such as about 2% polyacrylamide, preferably the polyacrylamide surface is coupled to a solid support such as glass. Microparticles and beads which facilitate reversible immobilization may be used. Solid phase reversible immobilization (SPRI) methods or modified methods are 20 known in the art and may be employed (e.g. see DeAngelis M. M. et al. (1995) SolidPhase Reversible Immobilization for the Isolation of PCR Products, Nucleic Acids Research, 23(22): 4742-4743.). Surfaces can be provided in the form of e.g. paramagnetic beads. Paramagnetic beads can agglomerate under the influence of a magnetic field. For example, 25 paramagnetic surfaces can be provided with chemical groups, e.g. carboxyl groups, which in appropriate attachment conditions will act as binding moieties for macromolecules including nucleic acids. Macromolecules can be eluted from such surfaces in appropriate elution conditions. Surfaces of microparticles and beads can be provided with UV-sensitive polycarbonate. Nucleic acids, for example, can be bound to 30 the activated surface in the presence of a suitable immobilization buffer. 2026234101   18 Sep 2026 Microparticles and beads may be allowed to move freely within a reaction solution and then reversibly immobilized, e.g. by holding the bead within a microwell or pit etched into a surface. A bead can be localised as part of an array e.g. by the use of a unique nucleic acid “barcode” attached to the bead or by the use of colour-coding. 5          The surface may be part of an electrowetting-on-dielectric system (EWOD). EWOD systems provide a dielectric-coated surface which facilitates microfluidic manipulation of very small liquid volumes in the form of microdroplets (e.g. see Chou, W-L., et al. (2015) Recent Advances in Applications of Droplet Microfluidics, Micromachines, 6: 1249-1271.). Droplet volumes can programmably be created, 10 moved, partitioned and combined on-chip by electrowetting techniques. Thus electrowetting systems provide alternative means to reversibly immobilize macromolecules to surfaces and / or to manipulate macromolecules immobilized to surfaces. Accordingly, in any one of the processes or uses according to the invention as 15   described or defined herein, the biochemical reaction may be performed in a solid phase reaction system which comprises a surface. In any one of the processes or uses according to the invention as described or defined herein wherein the biochemical reaction is performed in a solid phase reaction system which comprises a surface, any macromolecule required for the performance of 20 the reaction may be attached to the surface. For example, in one such process wherein the biochemical reaction is a process of amplifying a single stranded target nucleic acid molecule or a double stranded target nucleic acid molecule in the in vitro reaction system as described herein, the at least one nucleic acid primer, and / or the reaction macromolecule, and / or the IDR-macromolecule and / or the one or more polypeptide co- 25 factors may be attached to the surface. In any one of the processes or uses according to the invention as described or defined herein wherein the biochemical reaction is performed in a solid phase reaction system which comprises a surface, the IDR-macromolecule which is required for the performance of the reaction may be attached to the surface. 30          In any one of the processes or uses according to the invention as described or defined herein wherein the biochemical reaction is a recombinase polymerase 2026234101   18 Sep 2026 amplification process of amplifying a double stranded target nucleic acid molecule in the in vitro reaction system, wherein the reaction is performed in a solid phase reaction system which comprises a surface, and wherein the recombinase agent and / or the recombinase loading protein and / or the single strand stabilizing agent and / or the 5 polymerase and / or the exonuclease and / or the first nucleic acid primer and / or the second nucleic acid primer may be attached to the surface. In one such process or use, the first nucleic acid primer or the second nucleic acid primer may be attached to the surface. Alternatively, in other such processes or uses, both the first nucleic acid primer and the second nucleic acid primer may be attached to the surface. 10          In any one of the processes or uses according to the invention as described or defined herein wherein the biochemical reaction is performed in a solid phase reaction system, the surface to which a macromolecule is attached may be a microbead, preferably wherein the microbead comprises a silicon, glass, gel or a polymeric material, such as polystyrene, or any combination thereof. 15          In any one of the processes or uses described herein wherein the biochemical reaction is performed in a solid phase reaction system which comprises a surface and / or a substrate, the surface and / or the substrate may be provided as a flow-cell. Any suitable flow-cell compatible with the biochemical reaction being performed may be used. A suitable flow-cell may comprise a plurality of fluidic channels through which 20 reagents used to perform the biochemical reaction may flow. Any one or more macromolecules used to perform the biochemical reaction may be attached to a surface lining a fluidic channel. A suitable flow-cell may be used to perform a biochemical reaction for amplification of a single stranded or a double stranded target nucleic acid molecule. Sequencing reactions performed using the processes, uses and methods 25 described herein may also be performed using a suitable flow-cell. 2026234101   18 Sep 2026 EXAMPLES The following Examples are provided to illustrate the invention but not to limit the invention. 5 Example 1. Recombinase polymerase amplification of Listeria monocytogenes gene hly using Gp32 having an IDR tag derived from human Otxl. Purpose and summary of experiment 10 This experiment was performed to assess the performance of a Gp32 fusion protein preparation containing a tag comprising a histidine-rich amino acid domain sequence found in the intrinsically disordered region (IDR) of the human homeobox protein Otx 1. 15          The example demonstrates recombinase polymerase amplification (RPA) of the Listeria monocytogenes gene hly across a range of template concentrations using Gp32 C-terminally tagged with the histidine rich intrinsically disordered region (IDR) domain (Otxl) in the absence of a crowding agent. 20 Materials and methods The specific amino acid sequence of the IDR domain tag used was AGHHHHHPHAHHPLSQSSGHHHHHHHHHHQGYGGSG (SEQ ID NO:24). This was attached to the C-terminus of phage vB EcoM NBG1 Gp32. The recombinant 25 fusion protein was purified using standard 1-step immobilised metal (nickel) affinity chromatography relying on the histidines naturally present in the IDR domain tag of the fusion protein under test. The fusion protein was designated Gp32-HIS2. The full amino acid sequence of the fusion protein is presented as SEQ ID NO: 82 (Table 23). The recombinant phage vB EcoM NBG1 Gp32 fusion protein was then tested in 30 PEG-free amplification, i.e. in the absence of a crowding agent, using the indicated 2026234101   18 Sep 2026 copies of a DNA template derived from Listeria monocytogenes genomic DNA. The test template was titrated in copy number as indicated in Figure 1. Reactions were set up by mixing 25 mM Tris HC1 pH 8.3, 7.5 mM KOAc, 1 mM DTT, 2.5 mM ATP, 20 mM phosphocreatine, 1 pM creatine kinase, 1 mM dNTPs, 5   0.4 pM forward primer, 0.4 pM reverse primer, 0.12 pM probe, 20 pM Gp32 fusion, 4.8 pM UvsX, 8.6 pM UvsY, 0.135 pM 5. aureus DNA polymerase, and 0.27 pM Exonuclease III. Reactions were initiated by the addition of template at the given concentration and with 33 mM MgOAc. The relevant primers and probe are indicated below. 10       Forward primer: CGCCTGCAAGTCCTAAGACGCCAATCGAAAAGAAAC (SEQ ID NO:98). Reverse primer: CTGCATCTCCGTGGTATACTAATACATTGTTTTTA (SEQ ID NO:99). Probe: CGAAAAGAAACACGCGGATGAAATCGATAAG[FAM] 15 [THF][BHQ-1]ATACAAGGATTGGA (SEQ ID NO: 100), where FAM is fluorescein, THF is tetrahydrofuran and BHQ is Black Hole Quencher. Reactions were then incubated at 39°C and placed in a fluorometer with magnetic mixing using a bearing ball. 20 Results and conclusion As shown in Figure 1 the test template was readily detected at high sensitivity within 7 minutes of initiation of the RPA reaction. Amplicons were detected with as little as 10 copies of target. 25          It was therefore found that amplification in the absence of a crowding agent, such as PEG, occurred efficiently using this Gp32 IDR-tagged fusion protein. 2026234101   18 Sep 2026 Example 2. Recombinase polymerase amplification of the Listeria monocytogenes gene hly using Gp32 having an IDR tag derived from human MafA. Purpose and summary of experiment 5 This experiment was performed to assess the performance of a Gp32 fusion protein preparation containing a tag comprising a histidine-rich domain sequence found in the intrinsically disordered region (IDR) of the human transcription factor MafA. The example demonstrates recombinase polymerase amplification (RPA) of the 10 Listeria monocytogenes gene hly across a range of template concentrations using Gp32 C-terminally tagged with the histidine-rich intrinsically disordered region (IDR) domain (MafA) in the absence of a crowding agent. Materials and methods 15 The specific amino acid sequence of the IDR domain tag used was SGHHGAHHGAHHPAAAAAYEAFRGPGFAGGGGADDMGAGHHHGAHHAAHH HHAAHHHHHHHHHHGGAGHGGGAGHH (SEQ ID NO:27). This was attached to the C-terminus of phage vB EcoM NBG1 Gp32. The recombinant fusion protein was 20 purified using standard 1-step immobilised metal (nickel) affinity chromatography relying on the histidines naturally present in the IDR domain tag of the fusion protein under test. The fusion protein was designated Gp32-HIS5. The full amino acid sequence of the fusion protein is presented as SEQ ID NO: 85 (Table 23). The recombinant phage vB EcoM NBG1 Gp32 fusion protein was then tested in 25 PEG-free amplification, i.e. in the absence of a crowding agent, using the indicated copies of a DNA template derived from Listeria monocytogenes genomic DNA. The test template was titrated in copy number as indicated in Figure 2. Reactions were set up by mixing 25 mM Tris HC1 pH 8.3, 7.5 mM KOAc, 1 mM DTT, 2.5 mM ATP, 20 mM phosphocreatine, 1 pM creatine kinase, 1 mM dNTPs, 30   0.4 pM forward primer, 0.4 pM reverse primer, 0.12 pM probe, 20 pM Gp32 fusion, 4.8 pM UvsX, 8.6 pM UvsY, 0.135 pM S. aureus DNA polymerase, and 0.27 pM 2026234101   18 Sep 2026 Exonuclease III. Reactions were initiated by the addition of template at the given concentration and with 33 mM MgOAc. The relevant primers and probe are indicated below. Forward primer: CGCCTGCAAGTCCTAAGACGCCAATCGAAAAGAAAC 5 (SEQ ID NO:98). Reverse primer: CTGCATCTCCGTGGTATACTAATACATTGTTTTTA (SEQ ID NO:99). Probe: CGAAAAGAAACACGCGGATGAAATCGATAAG[FAM] [THF][BHQ-1]ATACAAGGATTGGA (SEQ ID NO: 100), where FAM is fluorescein, 10 THF is tetrahydro furan and BHQ is Black Hole Quencher. Reactions were then incubated at 39°C and placed in a fluorometer with magnetic mixing using a bearing ball. Results and conclusion 15 As shown in Figure 2 the test template was readily detected at high sensitivity within 10 minutes of initiation of the RPA reaction. Amplicons were detected with as little as 10 copies of target. It was therefore found that amplification in the absence of a crowding agent, 20 such as PEG, occurred efficiently using this Gp32 IDR-tagged fusion protein. Example 3. Recombinase polymerase amplification of the Listeria monocytogenes gene hly using Gp32 having an IDR tag derived from Saccharomyces cerevisiae Hrpl. 25 Purpose and summary of experiment This experiment was performed to assess the performance of a Gp32 fusion protein preparation containing a tag comprising an intrinsically disordered region (IDR) 30   of the Saccharomyces cerevisiae Hrpl protein. 2026234101   18 Sep 2026 The example demonstrates recombinase polymerase amplification (RPA) of the Listeria monocytogenes gene hly across a range of template concentrations using Gp32 C-terminally tagged with the sequence comprising an intrinsically disordered region (IDR) of the yeast Hrpl protein in the absence of a crowding agent. 5 Materials and methods The specific amino acid sequence of the IDR domain tag used was GGNNGGNNMNRRGGNFGNQGDFNQMYQNPMMGGYNPMMNPQAMTDYYQ 10 KMQEYYQQMQ (SEQ ID NO:9). This was attached to the C-terminus of phage vB EcoM NBG1 Gp32. The recombinant fusion protein was purified using standard 1-step immobilised metal (nickel) affinity chromatography relying on an additional heptahistidine tag placed at the very C-terminus of the fusion protein under test, i. e. placed after the IDR tag at the C-terminus of the fusion protein. The fusion protein was 15 designated Gp32-HRP1. The full amino acid sequence of the fusion protein is presented as SEQ ID NO: 79 (Table 23). The recombinant phage vB EcoM NBG1 Gp32 fusion protein was then tested in PEG-free amplification, i. e. in the absence of a crowding agent, using the indicated copies of a DNA template derived from Listeria monocytogenes genomic DNA. The 20 test template was titrated in copy number as indicated in Figure 3. Reactions were set up by mixing 25 mM Tris HC1 pH 8.3, 7.5 mM KOAc, 1 mM DTT, 2.5 mM ATP, 20 mM phosphocreatine, 1 pM creatine kinase, 1 mM dNTPs, 0.4 pM forward primer, 0.4 pM reverse primer, 0.12 pM probe, 20 pM Gp32 fusion, 4.8 pM UvsX, 8.6 pM UvsY, 0.135 pM S. aureus DNA polymerase, and 0.27 pM 25 Exonuclease III. Reactions were initiated by the addition of template at the given concentration and with 33 mM MgOAc. The relevant primers and probe are indicated below. Forward primer: CGCCTGCAAGTCCTAAGACGCCAATCGAAAAGAAAC (SEQ ID NO:98). 30       Reverse primer: CTGCATCTCCGTGGTATACTAATACATTGTTTTTA (SEQ ID NO :99). 2026234101   18 Sep 2026 Probe: CGAAAAGAAACACGCGGATGAAATCGATAAG[FAM] [THF][BHQ-1]ATACAAGGATTGGA (SEQ ID NO: 100), where FAM is fluorescein, THF is tetrahydrofuran and BHQ is Black Hole Quencher. Reactions were then incubated at 39°C and placed in a fluorometer with 5 magnetic mixing using a bearing ball. Results and conclusion As shown in Figure 3 the test template was readily detected at high sensitivity 10   within 7 minutes of initiation of the RPA reaction. Amplicons were detected with as little as 10 copies of target. It was therefore found that amplification in the absence of a crowding agent, such as PEG, occurred efficiently using this Gp32 IDR-tagged fusion protein. 15 Example 4. Recombinase polymerase amplification of the Listeria monocytogenes gene hly using Gp32 having an IDR tag derived from Saccharomyces cerevisiae Sup2. Purpose and summary of experiment 20 This experiment was performed to assess the performance of a Gp32 fusion protein preparation containing a tag comprising an intrinsically disordered region (IDR) domain of the Saccharomyces cerevisiae Sup2 protein. The example demonstrates recombinase polymerase amplification (RPA) of the 25 Listeria monocytogenes gene hly across a range of template concentrations using Gp32 C-terminally tagged with the sequence comprising an intrinsically disordered region (IDR) domain of the yeast Sup2 protein in the absence of a crowding agent. 2026234101   18 Sep 2026 Materials and methods The specific amino acid sequence of the IDR domain tag used was YNPQGGYQQ (SEQ ID NO:19). This was attached to the C-terminus of phage vB 5 EcoMNBGl Gp32. The recombinant fusion protein was purified using standard 1-step immobilised metal (nickel) affinity chromatography relying on an additional heptahistidine tag placed at the very C-terminus of the fusion protein under test, i. e. placed after the IDR domain tag at the C-terminus of the fusion protein. The fusion protein was designated Gp32-Supl. The full amino acid sequence of the fusion protein 10 is presented as SEQ ID NO: 72 (Table 23). The recombinant phage vB EcoM NBG1 Gp32 fusion protein was then tested in PEG-free amplification, i. e. in the absence of a crowding agent, using the indicated copies of a DNA template derived from Listeria monocytogenes genomic DNA. The test template was titrated in copy number as indicated in Figure C. 15         Reactions were set up by mixing 25 mM Tris HC1 pH 8.3, 7.5 mM KOAc, 1 mM DTT, 2.5 mM ATP, 20 mM phosphocreatine, 1 pM creatine kinase, 1 mM dNTPs, 0.4 pM forward primer, 0.4 pM reverse primer, 0.12 pM probe, 20 pM Gp32 fusion, 4.8 pM UvsX, 8.6 pM UvsY, 0.135 pM X aureus DNA polymerase, and 0.27 pM Exonuclease III. Reactions were initiated by the addition of template at the given 20 concentration and with 33 mM MgOAc. The relevant primers and probe are indicated below. Forward primer: CGCCTGCAAGTCCTAAGACGCCAATCGAAAAGAAAC (SEQ ID NO:98). Reverse primer: CTGCATCTCCGTGGTATACTAATACATTGTTTTTA (SEQ 25 ID NO:99). Probe: CGAAAAGAAACACGCGGATGAAATCGATAAG[FAM] [THF][BHQ-1]ATACAAGGATTGGA (SEQ ID NO: 100), where FAM is fluorescein, THF is tetrahydrofuran and BHQ is Black Hole Quencher. Reactions were then incubated at 39°C and placed in a fluorometer with 30 magnetic mixing using a bearing ball. 2026234101   18 Sep 2026 Results and conclusion As shown in Figure 4 the test template was readily detected at high sensitivity within 7 minutes of initiation of the RPA reaction. Amplicons were detected with as 5 little as 10 copies of target. It was therefore found that amplification in the absence of a crowding agent, such as PEG, occurred efficiently using this Gp32 IDR-tagged fusion protein. Example 5. Recombinase polymerase amplification of the human apoB gene using 10 Gp32 having IDR tags derived from Saccharomyces cerevisiae Sup2. Purpose and summary of experiment This experiment was performed to assess the performances of a number of Gp32 15   fusion protein preparations containing tags comprising an intrinsically disordered region (IDR) domain amino acid sequence of the Saccharomyces cerevisiae Sup2 protein. Variable numbers of an IDR domain repeat unit were assessed and a range of concentrations of the fusion proteins were examined. The example demonstrates recombinase polymerase amplification (RPA) of the 20 human apolipoprotein B (apoB) gene using Gp32 C-terminally tagged with the sequence comprising an intrinsically disordered region (IDR) domain of the yeast Sup2 protein in the absence of a crowding agent. Materials and methods 25 The specific amino acid sequence of the IDR domain tag used was YNPQGGYQQ (SEQ ID NO:19). This was attached to the C-terminus of phage vB EcoM NBG1 Gp32. Either a single YNPQGGYQQ unit was attached, or two, three or four repeats were attached. The recombinant fusion protein was purified using standard 30 1-step immobilised metal (nickel) affinity chromatography relying on an additional heptahistidine tag placed at the very C-terminus of the fusion protein under test, i. e. 2026234101   18 Sep 2026 placed after the IDR domain tag at the C-terminus of the fusion protein. The fusion proteins were designated Gp32-Sup2 (two repeats; SEQ ID NO:20), Gp32-Sup3 (three repeats; SEQ ID NO:21) and Gp32-Sup4 (four repeats; SEQ ID NO:22). The full amino acid sequences of the fusion proteins are presented respectively as SEQ ID NO: 5   73, SEQ ID NO: 74 and SEQ ID NO: 75 (Table 23). The recombinant phage vB EcoM NBG1 Gp32 fusion proteins were then tested, along with Gp32-Supl, in PEG-free amplification, i.e. in the absence of a crowding agent, using a DNA template derived from human genomic DNA. Reactions were set up by mixing 25 mM Tris HC1 pH 8.3, 7.5 mM KOAc, 1 10 mM DTT, 2.5 mM ATP, 20 mM phosphocreatine, 1 pM creatine kinase, 1 mM dNTPs, 0.4 pM forward primer, 0.4 pM reverse primer, 0.12 pM probe, Gp32 fusion protein at the concentrations indicated in Figures 5 A to D, 4.8 pM UvsX, 8.6 pM UvsY, 0.135 pM S. aureus DNA polymerase, and 0.27 pM Exonuclease III. Reactions were initiated by the addition of template and with 33 mM MgOAc. The test template copy number 15 used in each case was 10,000. The relevant primers and probe are indicated below. Forward primer: GCAGCTGTATAGCAAATTCCTGTTGAAAGCAG (SEQ ID NO:101). Reverse primer: TCCTGGCTGTATTCATTGTTGTTAAATTGG (SEQ ID 20 NO:102). Probe: CACTGATGCT TTTCCTAGACACGAGATGA[FAM-dT]G[THF]C[BHQl-dT]TGTGGAGCCTTTGT (SEQ ID NO: 103), where FAM is fluorescein, THF is tetrahydrofuran and BHQ is Black Hole Quencher. Reactions were then incubated at 39°C and placed in a fluorometer with 25 magnetic mixing using a bearing ball. Results and conclusion The results are shown in Figure 5. Figure 5A shows the results using a single 30 IDR domain tag unit. Figures 5 B to D show the results using respectively two, three 2026234101   18 Sep 2026 and four of the IDR domain tag unit repeats. The test template was detected after about 10 minutes of initiation of the RPA reaction. It was found that amplification in the absence of a crowding agent, such as PEG, occurred efficiently using these Gp32-IDR tagged fusion proteins. The best 5 performance was seen with a single IDR domain tag unit and with two IDR domain tag units. Three IDR domain tag units also gave good performance. Example 6. Recombinase polymerase amplification of the Listeria monocytogenes gene hly using Gp32 having an IDR tag derived from human MafA - comparison 10 of magnesium ion concentrations. Purpose of experiment This experiment was performed to assess the performance of a Gp32 fusion 15 protein preparation containing a tag comprising a histidine-rich domain sequence found in the intrinsically disordered region (IDR) of the human transcription factor MafA. The experiment assessed performance across a range of magnesium concentrations. The example demonstrates recombinase polymerase amplification (RPA) of the Listeria monocytogenes gene hly across a range of magnesium concentrations using 20 Gp32 C-terminally tagged with the histidine-rich intrinsically disordered region (IDR) domain (MafA) in the absence of a crowding agent. Materials and methods 25          The specific amino acid sequence of the IDR domain tag used was SGHHGAHHGAHHPAAAAAYEAFRGPGFAGGGGADDMGAGHHHGAHHAAHH HHAAHHHHHHHHHHGGAGHGGGAGHH (SEQ ID NO:27). This was attached to the C-terminus of phage vB EcoM NBG1 Gp32. The recombinant fusion protein was purified using standard 1 -step immobilised metal (nickel) affinity chromatography 30 relying on the histidines naturally present in the IDR domain tag of the fusion protein 2026234101   18 Sep 2026 under test. The fusion protein was designated Gp32-HIS5. The full amino acid sequence of the fusion protein is presented as SEQ ID NO: 85 (Table 23). The recombinant phage vB EcoM NBG1 Gp32 fusion protein was then tested in PEG-free amplification, i. e. in the absence of a crowding agent, using the indicated 5 copies of a DNA template derived from Listeria monocytogenes genomic DNA. The test template was provided at 10,000 copies per reaction and the magnesium ion concentration was varied from 5.6 mM up to 44.8 mM. Reactions were set up by mixing 25 mM Tris HC1 pH 8.3, 7.5 mM KOAc, 1 mM DTT, 2.5 mM ATP, 20 mM phosphocreatine, 1 pM creatine kinase, 1 mM dNTPs, 10   0.4 pM forward primer, 0.4 pM reverse primer, 0.12 pM probe, 20 pM Gp32 fusion, 4.8 pM UvsX, 8.6 pM UvsY, 0.135 pM S. aureus DNA polymerase, and 0.27 pM Exonuclease III. Reactions were initiated by the addition of template and the indicated concentration of MgOAc from 5.6 mM to 44.8 mM MgOAc. The relevant primers and probe are indicated below. 15       Forward primer: CGCCTGCAAGTCCTAAGACGCCAATCGAAAAGAAAC (SEQ ID NO:98). Reverse primer: CTGCATCTCCGTGGTATACTAATACATTGTTTTTA (SEQ ID NO :99). Probe: CGAAAAGAAACACGCGGATGAAATCGATAAG[FAM] 20 [THF][BHQ-1]ATACAAGGATTGGA (SEQ ID NO: 100), where FAM is fluorescein, THF is tetrahydrofuran and BHQ is Black Hole Quencher. Reactions were then incubated at 39°C and placed in a fluorometer with magnetic mixing using a bearing ball. 25 Results and conclusion It was found that amplification in the absence of a crowding agent such as PEG occurred efficiently using this Gp32 IDR-tagged fusion protein. As shown in Figure 6, it was found that good amplification occurred using this 30 Gp32 IDR-tagged fusion protein when 28 mM or more magnesium was present. The 2026234101   18 Sep 2026 optimum concentration in this experiment appeared to be 33.6 mM, and further increases up to 44.8 mM yielded similar time to detection. Example 7. Effects of phosphocreatine levels on recombinase polymerase 5 amplification of a human ApoB gene fragment. Purpose and summary of experiment This experiment was performed to assess the effects of varying the 10   phosphocreatine levels on the performance of a Gp32 fusion protein preparation containing a tag comprising a histidine-rich domain sequence found in the intrinsically disordered region (IDR) of the human homeobox protein Otxl. The example demonstrates recombinase polymerase amplification (RPA) of a fragment of the human apolipoprotein (apoB) gene using Gp32 C-terminally tagged 15 with the histidine-rich intrinsically disordered region (IDR) domain (Otxl) in the absence of a crowding agent. Materials and methods 20          The specific amino acid sequence of the IDR domain tag used was AGHHHHHPHAHHPLSQSSGHHHHHHHHHHQGYGGSG (SEQ ID NO:24). This was attached to the C-terminus of phage vB EcoM NBG1 Gp32. The recombinant fusion protein was purified using standard 1 -step immobilised metal (nickel) affinity chromatography relying on the histidines naturally present in the IDR domain tag of the 25 fusion protein under test. The fusion protein was designated Gp32-HIS2. The full amino acid sequence of the fusion protein is presented as SEQ ID NO: 82 (Table 23). The recombinant phage vB EcoM NBG1 Gp32 fusion protein was then tested in PEG-free amplification, i.e. in the absence of a crowding agent. A phosphocreatine titration was carried out using the human apoB assay. The test template was provided at 30 a concentration of 104 copies. 2026234101   18 Sep 2026 Reactions were set up by mixing 25 mM Tris HC1 pH 8.3, 7.5 mM KOAc, 1 mM DTT, 2.5 mM ATP, levels of phosphocreatine indicated in the figures, 1 pM creatine kinase, 1 mM dNTPs, 0.4 pM forward primer, 0.4 pM reverse primer, 0.12 pM probe, 20 pM Gp32 fusion, 4.8 pM UvsX, 8.6 pM UvsY, 0.135 pM S. aureus DNA 5 polymerase, and 0.27 pM Exonuclease III. Reactions were initiated by the addition of 104 copies template per reaction and with 33 mM MgOAc. Forward primer: GCAGCTGTATAGCAAATTCCTGTTGAAAGCAG (SEQ ID NO:101). Reverse primer: TCCTGGCTGTATTCATTGTTGTTAAATTGG (SEQ ID 10   NO:102). Probe: CACTGATGCT TTTCCTAGACACGAGATGA[FAM-dT]G[THF]C[BHQl-dT]TGTGGAGCCTTTGT (SEQ ID NO: 103), where FAM is fluorescein, THF is tetrahydrofuran and BHQ is Black Hole Quencher. Reactions were then incubated at 39°C and placed in a fluorometer with 15 magnetic mixing using a bearing ball. Results and conclusion It was found that amplification occured in the absence of a crowding agent such 20 as PEG using this Gp32 IDR-tagged fusion protein. As shown in Figures 7 A, B and C, at the standard phosphocreatine concentration used in PEG based RPA (50 mM), little amplification activity was seen within 20 minutes. Reducing phosphocreatine to 20 mM resulted in optimum performance, but good performance was also observed between 15-25 mM, and lower levels of amplification within 20 minutes was also 25 observed at 30-35 mM. 2026234101   18 Sep 2026 Example 8. Recombinase polymerase amplification of the Listeria monocytogenes gene hly using Gp32 having an IDR tag derived from Saccharomyces cerevisiae Hrpl - comparison of salt concentrations. 5 Purpose and summary of experiment This experiment was performed to assess the performance of a Gp32 fusion protein preparation containing a tag comprising an intrinsically disordered region (IDR) 10 of the Saccharomyces cerevisiae Hrpl protein. The experiment assessed performance across a range of salt concentrations, in this case using potassium acetate. The example demonstrates recombinase polymerase amplification (RPA) of the Listeria monocytogenes gene hly can be optimised across a range of salt concentrations using Gp32 C-terminally tagged with an intrinsically disordered region (IDR) of the 15   Saccharomyces cerevisiae Hrp 1 protein in the absence of a crowding agent. Materials and methods The specific amino acid sequence of the IDR domain tag used was 20 GGNNGGNNMNRRGGNFGNQGDFNQMYQNPMMGGYNPMMNPQAMTDYYQ KMQEYYQQMQ (SEQ ID NO:9). This was attached to the C-terminus of phage vB EcoM NBG1 Gp32. The recombinant fusion protein was purified using standard 1-step immobilised metal (nickel) affinity chromatography relying on an additional heptahistidine tag placed at the very C-terminus of the fusion protein under test, i. e. 25 placed after the IDR tag at the C-terminus of the fusion protein. The fusion protein was designated Gp32-HRP1. The full amino acid sequence of the fusion protein is presented as SEQ ID NO: 79 (Table 23). The recombinant phage vB EcoM NBG1 Gp32 fusion protein was then tested in PEG-free amplification, i. e. in the absence of a crowding agent, using 100 copies of a 30 DNA template derived from Listeria monocytogenes genomic DNA. The potassium acetate concentration was varied from 10 mM up to 100 mM. 2026234101   18 Sep 2026 Reactions were set up by mixing 25 mM Tris HC1 pH 8.3, 7.5 mM KOAc, 1 mM DTT, 2.5 mM ATP, 20 mM phosphocreatine, 1 pM creatine kinase, 1 mM dNTPs, 0.4 pM forward primer, 0.4 pM reverse primer, 0.12 pM probe, 20 pM Gp32 fusion, 4.8 pM UvsX, 8.6 pM UvsY, 0.135 pM S. aureus DNA polymerase, and 0.27 pM 5 Exonuclease III. Reactions were initiated by the addition of template and 33 mM MgOAc. The relevant primers and probe are indicated below. Forward primer: CGCCTGCAAGTCCTAAGACGCCAATCGAAAAGAAAC (SEQ ID NO:98). 10       Reverse primer: CTGCATCTCCGTGGTATACTAATACATTGTTTTTA (SEQ ID NO :99). Probe: CGAAAAGAAACACGCGGATGAAATCGATAAG[FAM] [THF][BHQ-1]ATACAAGGATTGGA (SEQ ID NO: 100), where FAM is fluorescein, THF is tetrahydrofuran and BHQ is Black Hole Quencher. 15          Reactions were then incubated at 39°C and placed in a fluorometer with magnetic mixing using a bearing ball. Results and conclusion 20          It was found that amplification in the absence of a crowding agent such as PEG occurred efficiently using this Gp32 IDR-tagged fusion protein. It was also found that amplification in the absence of a crowding agent using this Gp32 IDR-tagged fusion protein can be optimised across a range of salt conentrations, of which potassium acetate is a representative example. 25          As shown in Figure 8, it was found that good amplification occurred using this Gp32 IDR-tagged fusion protein when 10 mM or more potassium acetate was present. The optimum concentration range in this experiment appeared to be between 10 to 40 mM. At concentrations above 40 mM less efficient amplification was observed. 29040458.1:DCC-18 / 09 / 2026 2026234101   18 Sep 2026 Example 9. Recombinase polymerase amplification of a human ApoB gene fragment using Gp32 having an IDR tag derived from Saccharomyces cerevisiae Sup2 - synergistic effect with crowding agent. 5 Purpose and summary of experiment This experiment was performed to assess the effect of low concentrations of a crowding agent, in this case PEG, on the reaction efficiency of a Gp32 fusion protein preparation containing a histidine-rich sequence found in the intrinsically disordered 10 region of the yeast SUP2 gene, specifically the Sup1 sequence YNPQGGYQQ (SEQ ID NO:19), attached to the C-terminus of phage vB EcoM NBG1 Gp32. The performance of this fusion protein was compared to a Gp32 protein lacking the Sup1 IDR tag in recombinase polymerase amplification of a fragment of the human apolipoprotein (apoB) gene. 15          It was found that low concentrations of a crowding agent can enhance the reaction efficiency of the Sup1 IDR-tagged Gp32 and that conditions can be achieved where a synergistic effect can be observed. Materials and methods 20 Gp32-Sup1 The specific amino acid sequence of the IDR domain tag used was YNPQGGYQQ (SEQ ID NO:19). This was attached to the C-terminus of phage vB 25 EcoM NBG1 Gp32. The recombinant fusion protein was purified using standard 1-step immobilised metal (nickel) affinity chromatography relying on an additional heptahistidine tag placed at the very C-terminus of the fusion protein under test, i.e. placed after the IDR domain tag at the C-terminus of the fusion protein. The fusion protein was designated Gp32-Sup1. The full amino acid sequence of the fusion protein 30 is presented as SEQ ID NO: 72 (Table 23). 2026234101   18 Sep 2026 Gp32 (7His) Phage vB EcoM NBG1 Gp32 was purified using standard 1-step immobilised metal (nickel) affinity chromatography relying on a heptahistidine tag placed at the very 5 C-terminus of the protein under test. The fusion protein was designated Gp32 (7His). The full amino acid sequence of the fusion protein is presented as SEQ ID NO: 65 (Table 23). The recombinant phage vB EcoM NBG1 Gp32 fusion proteins were tested in RPA reactions, either in the presence or absence of a crowding agent using a DNA 10 template comprising a fragment of the human apolipoprotein (apoB) gene. Reactions were set up by mixing 25 mM Tris HC1 pH 8.3, 7.5 mM KOAc, 1 mM DTT, 2.5 mM ATP, 50 mM phosphocreatine, 1 pM creatine kinase, 1 mM dNTPs, 0.4 pM forward primer, 0.4 pM reverse primer, 0.12 pM probe, 20 pM Gp32 fusion protein, 4.8 pM UvsX, 8.6 pM UvsY, 0.135 pM S. aureus DNA polymerase, and 0.27 15 pM Exonuclease III. Reactions were initiated by the addition of template and with 33 mM MgOAc. The test template copy number used in each case was 10,000. PEG was added to the final concentrations noted in the relevant figure. The species of PEG used was PEG molecular weight 35,000. The relevant primers and probe are indicated below. 20       Forward primer: GCAGCTGTATAGCAAATTCCTGTTGAAAGCAG (SEQ ID NO:101). Reverse primer: TCCTGGCTGTATTCATTGTTGTTAAATTGG (SEQ ID NO:102). Probe: CACTGATGCT TTTCCTAGACACGAGATGA[FAM- 25 dT]G[THF]C[BHQl-dT]TGTGGAGCCTTTGT (SEQ ID NO: 103), where FAM is fluorescein, THF is tetrahydrofuran and BHQ is Black Hole Quencher. Reactions were then incubated at 39°C and placed in a fluorometer with magnetic mixing using a bearing ball. 2026234101   18 Sep 2026 Results and conclusion The results are shown in Figure 9. Figure 9 shows that when the Gp32-Supl 5 IDR-tagged fusion protein was tested in the absence of the crowding agent PEG the test template was detected efficiently. When the Gp32-7His fusion protein without the Supl IDR tag was tested in the presence of the crowding agent PEG, between 0.5% to 2%, a small but nevertheless detectable amount of amplified product was observed. 10          When the Gp32-Supl IDR-tagged fusion protein was tested in the presence of the crowding agent PEG the test template was detected efficiently. In this case a synergistic effect could be observed, with the amount of amplified product exceeding the combined amount when comparing the amount observed with: (i) the Gp32-Supl IDR-tagged fusion protein in the absence of PEG and (ii) the Gp32-7His fusion protein 15 without the Supl IDR tag in the absence of PEG (see for example Figure 9 and compare Supl 1% PEG with Supl 0% PEG + normal GP32 1% PEG). These results demonstrate that an enhanced effect on the performance efficiency of a biochemical reaction can be observed when combining an IDR-tagged macromolecular component of the reaction with low concentrations of a crowding 20 agent, and that conditions can be achieved that promote a synergistic effect on reaction efficiency when combining an IDR-tagged macromolecular component of the reaction with low concentrations of a crowding agent. Example 10. Promotion of phase separation by IDR tags in the presence of 25 multivalent metal cations. Purpose and summary of experiment This experiment was performed to assess, in an aqueous in vitro biochemical 30 system, the effects of multivalent metal cations on the promotion of phase separation 2026234101   18 Sep 2026 driven / caused by several Gp32 fusion proteins each having a tag comprising an intrinsically disordered region (IDR) domain amino acid sequence. The example demonstrates that tags comprising IDR domain amino acid sequence are surprisingly capable of promoting phase separation, and more surprisingly 5 that this effect is enhanced by the presence of multivalent metal cations. Materials and methods Gp32-HIS2 fusion protein 10 The specific amino acid sequence of the IDR domain tag used was AGHHHHHPHAHHPLSQSSGHHHHHHHHHHQGYGGSG (SEQ ID NO:24). This was attached to the C-terminus of phage vB EcoM NBG1 Gp32. The recombinant fusion protein was purified using standard 1 -step immobilised metal (nickel) affinity 15   chromatography relying on the histidines naturally present in the IDR domain tag of the fusion protein under test. The full amino acid sequence of the Gp32-HIS2 fusion protein is provided as SEQ ID NO:82 (Table 23). Gp32-HRP1 fusion protein 20 The specific amino acid sequence of the IDR domain tag used was GGNNGGNNMNRRGGNFGNQGDFNQMYQNPMMGGYNPMMNPQAMTDYYQ KMQEYYQQMQ (SEQ ID NO:9). This was attached to the C-terminus of phage vB EcoM NBG1 Gp32. The recombinant fusion protein was purified using standard 1-step 25 immobilised metal (nickel) affinity chromatography relying on an additional heptahistidine tag placed at the very C-terminus of the fusion protein under test, i. e. placed after the IDR tag at the C-terminus of the fusion protein. The full amino acid sequence of the Gp32-HRP1 fusion protein is provided as SEQ ID NO:79 (Table 23). 2026234101   18 Sep 2026 Gp32-Supl fusion protein The specific amino acid sequence of the IDR domain tag used was 5 YNPQGGYQQ (SEQ ID NO:19). This was attached to the C-terminus of phage vB EcoM NBG1 Gp32. The recombinant fusion protein was purified using standard 1-step immobilised metal (nickel) affinity chromatography relying on an additional heptahistidine tag placed at the very C-terminus of the fusion protein under test, i. e. placed after the IDR domain tag at the C-terminus of the fusion protein. The full amino 10 acid sequence of the Gp32-Supl fusion protein is provided as SEQ ID NO:72 (Table 23). Gp32-Fib fusion protein 15          The specific amino acid sequence of the IDR domain tag used was PGFSPRGGGFGGRGGFGDRGGRGGRGGFGGGRGRGGGFRGRGR (SEQ ID NO:1). This was attached to the C-terminus of phage vB EcoM NBG1 Gp32. The recombinant fusion protein was purified using standard 1 -step immobilised metal (nickel) affinity chromatography relying on an additional heptahistidine tag placed at 20 the very C-terminus of the fusion protein under test, i.e. placed after the IDR domain tag at the C-terminus of the fusion protein. The full amino acid sequence of the Gp32-Fib fusion protein is provided as SEQ ID NO:69 (Table 23). Phase separation assay 25 The methods outlined below apply for all the fusion proteins tested. The volumes of the fusion protein solutions used depended upon the protein concentrations after purification. A 50 pl solution was made up in each case, comprising the tagged fusion protein 30 at a final concentration of 1000 ng / pl and metal ion, in either the acetate or the chloride 2026234101   18 Sep 2026 form, at the target concentration indicated below and in the relevant figures presented herein. For the Gp32-HIS2 fusion, protein concentration after purification was 48 mg / ml. 1.04 pl of this fusion protein was used in each 50 pl reaction to achieve a final 5 concentration of 1000 ng / pl in solution. For the Gp32-HRP1 fusion, protein concentration after purification was 39 mg / ml. 1.28 pl of this fusion protein was used in each 50 pl reaction to achieve a final concentration of 1000 ng / pl in solution. For the Gp32-Supl fusion, protein concentration after purification was 36 mg / ml. 1.4 pl of this fusion protein was used in each 50 pl reaction to achieve a final concentration of 1000 10 ng / pl in solution. For the Gp32-Fib fusion, protein concentration after purification was 20.2 mg / ml. 2.48 pl of this fusion protein was used in each 50 pl reaction to achieve a final concentration of 1000 ng / pl in solution. In these experiments the divalent metal cation concentration required for detectible phase separation enhancement to occur was tested with representative 15 divalent metal cations: magnesium (MgOAc), manganese (MgCF) and calcium (CaCh). The acetate forms of manganese and calcium were not used simply due to their known instability in solution. Manganese will oxidise over time in the acetate solution, and calcium acetate seems to support the growth of some bacteria in solution whereas calcium chloride does not. 20          Following constitution of a mixture comprising water, the IDR-tagged protein and divalent metal cations, the mixture was vortexed, spun down and a 10 pl sample of the mixture was transferred to a DHC-B01 C-Chip haemocytometer slide. The slide was then imaged under a microscope at x400 magnification. Detectibe phase separation was assessed by the formation of spherical-like globular foci / particles which can be 25 identified visually via magnification and counted using the haemocytometer. A globular foci count per unit volume can then be performed. Globular foci counts were performed by counting the number of globular foci which formed in a magnification area of 218 pm x 175 pm at x400 magnification. This was done by splitting the magnified image into 20 square segments (4 x 5 of the image), counting the globular 30 foci in one of these segments and then multiplying this number by 20. 2026234101   18 Sep 2026 Results and conclusions It was observed that the transition that occurs between just below the minimum 5 detectible phase separating concentration (MPSC) in this assay and just above the MPSC occurred very suddenly in all of the reactions performed. At just below the MPSC no detectible phase-separated aqueous particles were observed at all, and the solution was found to be empty of visuably detectible particles (globular foci). Above the MPSC the transition was very apparent with hundreds of visuably detectible 10 particles (globular foci) suddenly forming. The size of the globular foci varied and was found to be corellated with the IDR tag and the divalent metal cation used. It was determined that the specific sizes of the globular foci was unimportant. Globular foci were present as particle-like structures being broadly spherical in 15 shape. For any given IDR tag and any given divalent metal ion combination an average diameter of a population of globular foci can readily be determined using standard methods. Results using individual fusion proteins are outlined below. 20 Gp32-HIS2 fusion protein The minimum concentration of magnesium required to enhance the formation of detectible phase-separated aqueous particles in these conditions was determined to be 10 mM - approximately 600 particles (globular foci) were counted within the field. 25          The minimum concentration of calcium ions required to enhance the formation of detectible phase-separated aqueous particles in these conditions was determined to be 12 mM - approximately 500 particles (globular foci) were counted within the field. The minimum concentration of manganese ions required to enhance the formation of detectible phase-separated aqueous particles in these conditions was 30 determined to be 2 mM - approximately 180 particles (globular foci) were counted within the field. 2026234101   18 Sep 2026 Representative magnified images are shown in Figure 10A. Gp32-HRP1 fusion protein 5          The minimum concentration of magnesium ions required to enhance the formation of detectible phase-separated aqueous particles in these conditions was determined to be 16 mM. At this concentration approximately 580 particles (globular foci) were counted within the field. The minimum concentration of calcium ions required to enhance the formation 10 of detectible phase-separated aqueous particles in these conditions was determined to be 24 mM. At this concentration approximately 240 particles (globular foci) were counted within the field. The minimum concentration of manganese ions required to enhance the formation of detectible phase-separated aqueous particles in these conditions was 15 determined to be 6 mM. At this concentration approximately 260 particles (globular foci) were counted within the field. Representative magnified images are shown in Figure 10B. Gp32-Supl fusion protein 20 The minimum concentration of magnesium ions required to enhance the formation of detectible phase-separated aqueous particles in these conditions was determined to be 24 mM. At this concentration approximately 280 particles (globular foci) were counted within the field. 25          The minimum concentration of calcium ions required to enhance the formation of detectible phase-separated aqueous particles in these conditions was determined to be 32 mM. At this concentration approximately 460 particles (globular foci) were counted within the field. The minimum concentration of manganese ions required to enhance the 30 formation of detectible phase-separated aqueous particles in these conditions was 2026234101   18 Sep 2026 determined to be 4 mM. At this concentration approximately 220 particles (globular foci) were counted within the field. Representative magnified images are shown in Figure 10C. 5 Gp32-Fib fusion protein The minimum concentration of magnesium ions required to enhance the formation of detectible phase-separated aqueous particles in these conditions was determined to be 500 pM. At this concentration approximately 340 particles (globular 10   foci) were counted within the field. The minimum concentration of calcium ions required to enhance the formation of detectible phase-separated aqueous particles in these conditions was determined to be 1 mM. At this concentration approximately 500 particles (globular foci) were counted within the field. 15          The minimum concentration of manganese ions required to enhance the formation of detectible phase-separated aqueous particles in these conditions was determined to be 500 pM. At this concentration approximately 360 particles (globular foci) were counted within the field. Representative magnified images are shown in Figure 10D. 20          Using these assays it was determined that the functional capability of an IDR or IDR domain to enhance the formation of detectible phase-separated aqueous particles in an in vitro biochemical environment when tagged to a protein can be established when 10 or more particles (globular foci) were are formed in a magnification area of 218 pm x 175 pm at x400 magnification. The functional capability of an IDR or IDR domain to 25 induce phase separation in an in vitro biochemical environment when tagged to a protein can be established when preferably 50 or more particles (globular foci) were are formed in a magnification area of 218 pm x 175 pm at x400 magnification, more preferably when 100 or more particles (globular foci) were are formed. The term “globular foci” as used herein is synonymous with “globule”, 30 “particle” or “globular particle” and these terms can be used interchangeably. 2026234101   18 Sep 2026 Example 11. Formation of globular foci by IDR tags in the presence of multivalent metal cations. Purpose and summary of experiment 5 This experiment was performed to assess, in an in vitro biochemical reaction system, the effects of multivalent metal cations on the promotion of phase separation driven / caused by several Gp32 fusion proteins each having a tag comprising an intrinsically disordered region (IDR) domain amino acid sequence. 10          The example demonstrates that tags comprising IDR domain amino acid sequences are capable of promoting / enhancing phase separation, and that this effect occurs in the presence of various multivalent m...

Claims

1. A process of performing a biochemical reaction in an aqueous in vitro reaction system, wherein the biochemical reaction is dependent on the function of at least one5 reaction macromolecule, optionally at least one reaction polypeptide, the process comprising: introducing at least one IDR-macromolecule into the in vitro reaction system under conditions suitable for performing the reaction, wherein the at least one IDR-macromolecule comprises one or more functional intrinsically disordered regions (IDRs), wherein upon introduction of the at least one IDR-macromolecule into the in10   vitro reaction system the efficiency of the biochemical reaction is increased by the atleast one IDR-macromolecule; preferably wherein the at least one IDR-macromolecule is at least one IDR-polypeptide.

2. A process according to claim 1, wherein the biochemical reaction is dependent15 on the function of the at least one IDR-macromolecule, optionally the at least one IDR-polypeptide, wherein upon introduction into the system the at least one IDR-macromolecule or the at least one IDR-polypeptide performs its reaction function in the biochemical reaction and increases the efficiency of the reaction in the system.20   3. A process according to claim 1 or claim 2, further comprising maintaining theIDR-macromolecule or the IDR-polypeptide in the system to cause liquid-liquid demixing and the formation of a plurality of phase-separated aqueous compartments within the system by the IDR-macromolecule or the IDR-polypeptide, thereby increasing the efficiency of the biochemical reaction in the system.

254. A process according to claim 3, further comprising maintaining the IDR-macromolecule or the IDR-polypeptide in the system to cause molecules necessary for the performance of the reaction to co-localise with the IDR-macromolecule or the IDR-polypeptide within the plurality of phase-separated aqueous compartments, thereby30 increasing the efficiency of the biochemical reaction in the system.2026234101   18 Sep 20265. A process according to claim 3 or claim 4, wherein the plurality of phase-separated aqueous compartments is a plurality of detectable phase-separated aqueous particles.5   6. A process according to any one of the preceding claims, wherein the process is abiochemical reaction for synthesizing a nucleic acid molecule in an in vitro reaction system comprising:(a) providing at least one nucleic acid primer;(b) providing a target nucleic acid molecule comprising at least one target10               strand, and contacting the at least one nucleic acid primer with the targetstrand, thereby forming a double stranded structure;(c) providing the IDR-macromolecule as an IDR-polypeptide, wherein the IDR-polypeptide is polymerase or one or more polypeptide co-factors; and(d) allowing the reaction to proceed, thereby extending the 3 ’ end of the at least15              one nucleic acid primer with polymerase and dNTPs, optionally in thepresence of one or more polypeptide co-factors, to generate a double stranded nucleic acid, wherein the first strand comprises a sequence of the target strand and the second strand comprises a sequence which is complementary thereto.

207. A process according to claim 6, wherein the process is a biochemical reaction for amplifying a single stranded target nucleic acid molecule or a double stranded target nucleic acid molecule in an in vitro reaction system, preferably wherein the target nucleic acid molecule is a DNA molecule.

258. A process according to any one of claims 1 to 5, wherein the process is a biochemical reaction for amplifying a double stranded target nucleic acid molecule in an in vitro reaction system comprising:(a) providing first and second nucleic acid primers;30          (b) providing a double stranded target nucleic acid molecule comprising a firststrand and a second strand, and contacting the first and second nucleic acid2026234101   18 Sep 2026primers with the target nucleic acid molecule thereby forming a first double stranded structure with the first strand and a second double strandedstructure with the second strand;(c) providing the IDR-macromolecule as an IDR-polypeptide, wherein the IDR-5              polypeptide is polymerase or one or more protein co-factors;(d) allowing the reaction to proceed, thereby extending the 3 ’ ends of the first and second nucleic acid primers with polymerase and dNTPs, optionally in the presence of one or more protein co-factors, to generate first and second double stranded nucleic acids; and10           (e) repeating steps (b) to (d) until a desired degree of amplification is reached.

9. A process according to claim 8, wherein the process is a recombinase polymerase amplification process of amplifying the double stranded target nucleic acid molecule in the in vitro reaction system comprising:15          (a) providing a recombinase agent, optionally a recombinase loading protein, asingle strand stabilizing agent, polymerase, first and a second nucleic acid primers, double stranded target nucleic acid comprising a first strand and a second strand, and optionally an exonuclease such as Exonuclease III;(b) contacting the recombinase agent with the first and second nucleic acid20              primers and optionally with the recombinase loading protein to form firstand second nucleoprotein primers which comprise a single stranded region at their 3 ’ ends;(c) contacting the first and second nucleoprotein primers with the target nucleic acid molecule thereby forming a first double stranded structure with the first25              strand and a second double stranded structure with the second strand;(d) allowing the reaction to proceed, thereby extending the 3 ’ end of the first and second nucleoprotein primers with polymerase and dNTPs to generate first and second double stranded nucleic acids and first and a second displaced nucleic acid strands, wherein the single strand stabilizing agent30               stabilizes the first and second displaced strands; and2026234101   18 Sep 2026(e) continuing the reaction by repeating steps (b) to (d) until a desired degree of amplification is reached;wherein the recombinase agent, and / or the recombinase loading protein, and / or the single strand stabilizing agent, and / or the polymerase is provided as the IDR-5 polypeptide.

10. A process according to claim 9, wherein the recombinase agent is selected from the group consisting of UvsX, T4 UvsX, T6 UvsX, RB18 UvsX, E.coli phage wV7 UvsX, Shigella phage CB8 UvsX, Shigella phage Shfl2 UvsX, E.coli phage ARI UvsX, 10 phage vB_EcoM_G4507 UvsX, Shigella phage SHFML-11 UvsX, Escherichia phagevBEcoMDalCa UvsX, E. coli RecA, E. coli RadA, E. coli RadB, E. coli Rad 51 or any functional analog, homolog or derivative thereof, and any combination thereof, preferably wherein the recombinase agent is UvsX, more preferably Escherichia phage vBEcoMDalCa UvsX.1511. A process according to claim 9 or claim 10, wherein the process includes a recombinase loading protein and wherein the recombinase loading protein is selected from the group consisting of UvsY, E. coli RecO, E. coli RecR or any functional analog, homolog or derivative thereof, and any combination thereof, preferably wherein20 the recombinase loading protein is UvsY, more preferably Escherichia phage STO UvsY.

12. A process according to any one of claims 6 to 11, wherein the polymerase is a eukaryotic polymerase selected from the group consisting of pol-a, pol-0, pol-6, pol-e or 25 any functional analog, homolog or derivative thereof, and any combination thereof.

13. A process according to any one of claims 6 to 11, wherein the polymerase is a prokaryotic polymerase selected from the group consisting of Bacillusstearothermophilus polymerase I large fragment, Bacillus subtilis Pol I large fragment 30 (Bsu polymerase), Listeria monocytogenes DNA polymerase I, S. aureus DNApolymerase I (Sau polymerase), E. coli DNA polymerase I Klenow fragment, E. coli2026234101   18 Sep 2026DNA polymerase I, E. coli DNA polymerase II, E. coli DNA polymerase III, E. coli DNA polymerase IV, E. coli DNA polymerase V, or any functional analog, homolog or derivative thereof, and any combination thereof, preferably wherein the polymerase is S. aureus DNA polymerase I (Sau polymerase) or Bacillus subtilis Pol I large fragment5 (Bsu polymerase).

14. A process according to any one of claims 6 to 11, wherein the polymerase is a bacteriophage polymerase selected from the group consisting of bacteriophage T4 gp43 DNA polymerase, T7 DNA polymerase and Phi-29 DNA polymerase, or any functional10 analog, homolog or derivative thereof, and any combination thereof.

15. A process according to any one of claims 9 to 14, wherein the single strand stabilizing agent is selected from the group consisting of Gp32, E. coli SSB protein, phage T4 Gp32 protein, phage Rb69 Gp32, phage vB_EcoM_NBGl Gp32, or any15 functional analog, homolog or derivative thereof, and any combination thereof, preferably the single strand stabilizing agent is Gp32 or phage vB_EcoM_NBGl Gp32.

16. A process according to any one of claims 9 to 15, wherein only the recombinase agent is provided as the IDR-polypeptide, or wherein only the recombinase loading20 protein is provided as the IDR-polypeptide, or wherein only the single strand stabilizing agent is provided as the IDR-polypeptide, or wherein only the polymerase is provided as the IDR-polypeptide, or wherein only the exonuclease is provided as the IDR-polypeptide.25   17. A process according to any one of the preceding claims, wherein the one ormore functional IDRs of the IDR-polypeptide are tagged to the IDR-polypeptide as an amino acid sequence comprising or consisting of the one or more IDRs such that the IDR-polypeptide is a genetically engineered fusion protein, wherein the one or more functional IDRs are located at the C-terminus of the IDR-polypeptide, at the N-terminus30 of the IDR-polypeptide, or at both the C-terminus of the IDR-polypeptide and the N-2026234101   18 Sep 2026terminus of the IDR-polypeptide, or at any amino acid position along the length of the polypeptide.

18. A process according to any one of the preceding claims, wherein the one or5 more functional IDRs of the IDR-macromolecule or the IDR-polypeptide is characterised as a sequence of amino acids which scores greater than 0.5 when analysed by the algorithm MetaDisorder.

19. A process according to any one of the preceding claims, wherein the one or10   more functional IDRs of the IDR-macromolecule or the IDR-polypeptide comprises orconsists of an amino acid sequence which includes one or more repeats of the tripeptide sequence RGG.

20. A process according to claim 19, wherein the one or more functional IDRs of the 15   IDR-macromolecule or the IDR-polypeptide comprises or consists of an amino acidsequence which further includes one or more repeats of the di-peptide sequence FG.

21. A process according to claim 19 or claim 20, wherein the one or more functional IDRs of the IDR-macromolecule or the IDR-polypeptide comprises or consists of an 20 amino acid sequence which further includes at least one aromatic amino acid residue consisting of tyrosine or phenylalanine.

22. A process according to any one of the preceding claims, wherein the one or more functional IDRs of the IDR-macromolecule or the IDR-polypeptide comprises or 25 consists of the amino acid sequence ofi. (YNPQGGYQQ)n, wherein n is a positive integer between 1 and 10, optionally wherein n = 1, 2, or 3; orii. (YSPTSPS)n, wherein n is a positive integer between 1 and 10, optionally wherein n = 1, 2, or 3; or30 iii. (FSPTSPT)n, wherein n is a positive integer between 1 and 10, optionally wherein n = 1, 2, or 3; or2026234101   18 Sep 2026iv. (YSPTSP-A / N / G)n, wherein n is a positive integer between 1 and 10, optionally wherein n = 1, 2, or 3; orv. (YSPGSPA)n, wherein n is a positive integer between 1 and 10, optionally wherein n = 1, 2, or 3.

523. A process according to any one of the preceding claims, wherein the one or more functional IDRs of the IDR-macromolecule or the IDR-polypeptide comprises or consists of an amino acid sequence which is glutamine rich, optionally wherein the amino acid sequence comprises at least 4, at least 5, at least 6, at least 7, at least 8, at10 least 9, or at least 10 consecutive glutamine residues.

24. A process according to claim 23, wherein the one or more functional IDRs of the IDR-macromolecule or the IDR-polypeptide comprises or consists of an amino acid sequence comprising one or more repeats of the tri-peptide sequence QQQ.1525. A process according to claim 23 or claim 24, wherein the one or more functional IDRs of the IDR-macromolecule or the IDR-polypeptide comprises or consists of the amino acid sequence of (QQQPQY)n, wherein n is a positive integer between 1 and 10, optionally wherein n = 1, 2, or 3.2026. A process according to any one of the preceding claims, wherein the one or more functional IDRs of the IDR-macromolecule or the IDR-polypeptide comprises or consists of a sequence of at least 5 consecutive amino acids of SEQ ID NO: 1.25   27. A process according to any one of the preceding claims, wherein the one ormore functional IDRs of the IDR-macromolecule or the IDR-polypeptide comprises or consists of an amino acid sequence of at least 5 consecutive amino acids of SEQ ID NO:9.30   28. A process according to any one of the preceding claims, wherein the one ormore functional IDRs of the IDR-macromolecule or the IDR-polypeptide comprises an2026234101   18 Sep 2026amino acid sequence containing one or more aromatic tyrosine residues and one or more phenylalanine residues which can engage in aromatic cation-pi interactions with multivalent metal ions, preferably divalent metal ions.5   29. A process according to any one of the preceding claims, wherein the one ormore functional IDRs of the IDR-macromolecule or the IDR-polypeptide comprises an amino acid sequence containing one or more arginine residues which can engage in guanidine-metal interactions with multivalent metal ions, preferably divalent metal ions.10   30. A process according to any one of the preceding claims, wherein the IDR-polypeptide comprises or consists of the polypeptide tagged with an amino acid sequence which comprises or consists of the amino acid sequence of any one of SEQ ID NOs 1 to 43, or which comprises or consists of a functional variant amino acid sequence of SEQ ID NOs 1 to 43, e.g. which has 80% or more identity to any one of SEQ ID NOs15 Ito 43.

31. A process according to any one of claims 9 to 30, wherein the IDR-polypeptide is a single strand stabilizing agent which is Gp32 and which has the amino acid sequence of any one of SEQ ID NOs 65 to 88, or wherein the IDR-polypeptide is a20 functional variant thereof, e.g. an IDR-polypeptide having an amino acid sequence which has 80% or more identity to any one of SEQ ID NOs 65 to 88.

32. A process according to any one of claims 9 to 30, wherein the IDR-polypeptide is a recombinase agent which is UvsX and which has the amino acid sequence of any25 one of SEQ ID NOs 44 to 59, or wherein the IDR-polypeptide is a functional variant thereof, e.g. an IDR-polypeptide having an amino acid sequence which has 80% or more identity to any one of SEQ ID NOs 44 to 59.

33. A process according to any one of claims 9 to 30, wherein the IDR-polypeptide30 is a recombinase loading protein which is UvsY and which has the amino acid sequence of any one of SEQ ID NOs 60 to 64, or wherein the IDR-polypeptide is a functional2026234101   18 Sep 2026variant thereof, e.g. an IDR-polypeptide having an amino acid sequence which has 80% or more identity to any one of SEQ ID NOs 60 to 64.

34. A process according to any one of the preceding claims, further comprising5 providing multivalent metal ions to the IDR-macromolecule or IDR-polypeptide in the in vitro reaction system, thereby simulating or enhancing the liquid-liquid demixing in the in vitro reaction system, thereby increasing the efficiency of the biochemical reaction, preferably wherein said liquid-liquid demixing promotes the formation of detectable phase-separated particles in the in vitro reaction system thereby increasing10   the efficiency of the biochemical reaction; optionally wherein the multivalent metal ionsare provided at a concentration of about 22 mM or more, preferably wherein the multivalent metal ions are provided at a concentration of between about 22 mM to 50 mM.15   35. A process according to claim 34, wherein the multivalent metal ions are divalentmetal ions, optionally Mg2+, Mn2+, Ca2+, Co2+ or Ni2+, preferably Mg2+, Mn2+ or Ca2+, more preferably Mg2+.

36. A process according to any one of the preceding claims, wherein the20 biochemical reaction is performed in a solid phase reaction system which comprises a surface.

37. A process according to claim 36, wherein the biochemical reaction is a process of amplifying a single stranded target nucleic acid molecule or a double stranded target 25 nucleic acid molecule in the in vitro reaction system as defined in claim 3 or claim 4, wherein the at least one nucleic acid primer and / or the IDR-macromolecule and / or the one or more polypeptide co-factors are attached to the surface.

38. A process according to any one of claims 9 to 34, wherein the biochemical30 reaction is a recombinase polymerase amplification process of amplifying a double stranded target nucleic acid molecule in the in vitro reaction system, wherein the2026234101   18 Sep 2026reaction is performed in a solid phase reaction system which comprises a surface, and wherein the recombinase agent and / or the recombinase loading protein and / or the single strand stabilizing agent and / or the polymerase and / or the exonuclease and / or the first nucleic acid primer and / or the second nucleic acid primer is / are attached to the surface,5 preferably wherein: (i) the first nucleic acid primer or the second nucleic acid primer is attached to the surface; or (ii) both the first and second nucleic acid primers are attached to the surface.

39. A process according to any one of claims 36 to 38, wherein the surface is planar 10 or is a microbead, preferably wherein the surface comprises silicon, glass, a gel-basedmaterial and / or a polymeric material such as polystyrene, more preferably wherein the surface is a microbead comprising a polymeric material such as polystyrene.

40. A process according to claim 39, wherein the surface is bound to a substrate,15   preferably wherein the surface is planar and / or the substrate comprises glass, optionallywherein the surface and / or the substrate are provided as a flow-cell.

41. A non-naturally occurring IDR-macromolecule comprising a macromolecule and a tag amino acid sequence, wherein the tag sequence comprises or consists of one20 or more functional intrinsically disordered regions (IDRs), wherein the IDR-macromolecule is capable of promoting liquid-liquid demixing in solution.

42. An IDR-macromolecule according to claim 41, wherein said liquid-liquid demixing promotes the formation of detectable phase-separated particles in the in vitro25 reaction system.

43. An IDR-macromolecule according to claim 41 or claim 42, wherein said liquidliquid demixing and formation of phase-separated aqueous compartments thereby increases the efficiency of the biochemical reaction in the system, or wherein the30   formation of the detectable phase-separated aqueous particles thereby increases theefficiency of the biochemical reaction in the system.2026234101   18 Sep 202644. An IDR-macromolecule according to any one of claims 41 to 43, which is a non-naturally occurring, artificial or genetically engineered IDR-polypeptide comprising a polypeptide and the tag amino acid sequence.

545. An IDR-polypeptide according to claim 44, wherein the tag amino acid sequence is located at the C-terminus of the polypeptide, at the N-terminus of the polypeptide or at both the C-terminus of the polypeptide and the N-terminus of the polypeptide, or at any amino acid position along the length of the polypeptide.1046. An IDR-polypeptide according to claim 45, wherein the one or more functional IDRs of the tag amino acid sequence are functional IDRs as defined in any one of claims 14 to 28.15   47. An IDR-macromolecule or polypeptide according to any one of claims 41 to 46,wherein the tag sequence comprises amino acid residues which are capable of engaging in aromatic cation-pi interactions with multivalent metal cations, preferably divalent metal cations, more preferably Mg2+, Mn2+, Ca2+, Co2+ or Ni2+ ions, yet more preferably Mg2+, Mn2+ or Ca2+, even more preferably Mg2+.2048. An IDR-polypeptide according to any one of claims 44 to 47, wherein the polypeptide to which the sequence comprising or consisting of the one or more functional IDRs is tagged is an enzyme, such as a helicase, a gyrase, a recombinase, e.g. an RPA recombinase agent, a nuclease, e.g. an exonuclease and an endonuclease, a25 ligase, a glycolyase, a methylase, a methyltransferase, a glucosyltransferase, a polymerase, a kinase, a phosphatase, a gene editing enzyme such as a CRISPR enzyme, e.g. a Cas9 enzyme; a co-factor, e.g. as an RPA recombinase loading protein and an RPA single strand stabilizing agent.30   49. An IDR-polypeptide according to claim 48, wherein the polypeptide to whichthe sequence comprising or consisting of the one or more functional IDRs is tagged is2026234101   18 Sep 2026an RPA single strand stabilizing agent, preferably Gp32; optionally wherein the IDR-polypeptide has the amino acid sequence of any one of SEQ ID NOs 65 to 88, or wherein the IDR-polypeptide is a functional variant thereof, e.g. an IDR-polypeptide having an amino acid sequence which has 80% or more identity to any one of SEQ ID 5 NOs 65 to 88.

50. An IDR-polypeptide according to claim 48, wherein the polypeptide to which the sequence comprising or consisting of the one or more functional IDRs is tagged is an RPA recombinase agent, preferably UvsX; optionally wherein the IDR-polypeptide 10 has the amino acid sequence of any one of SEQ ID NOs 44 to 59, or wherein the IDR-polypeptide is a functional variant thereof, e.g. an IDR-polypeptide having an amino acid sequence which has 80% or more identity to any one of SEQ ID NOs 44 to 59.

51. An IDR-polypeptide according to claim 48, wherein the polypeptide to which 15 the sequence comprising or consisting of the one or more functional IDRs is tagged isan RPA recombinase loading protein, preferably UvsY; optionally wherein the IDR-polypeptide has the amino acid sequence of any one of SEQ ID NOs 60 to 64, or wherein the IDR-polypeptide is a functional variant thereof, e.g. an IDR-polypeptide having an amino acid sequence which has 80% or more identity to any one of SEQ ID 20 NOs 60 to 64.

52. An isolated nucleic acid molecule comprising a first nucleic acid sequence encoding the IDR-polypeptide according to any one of claims 41 to 51; optionally including a second nucleic acid sequence encoding a promoter, wherein the first nucleic 25 acid sequence is operably linked to the second nucleic acid sequence.

53. A recombinant polynucleotide expression vector comprising the nucleic acid molecule according to claim 52.30   54. A host cell comprising the nucleic acid molecule defined in claim 52, or therecombinant polynucleotide expression vector defined in claim 53.2026234101   18 Sep 202655. A cell culture comprising a growth medium and a population of host cells, wherein the population comprises host cells as defined according to claim 54.5   56. A kit comprising a non-naturally occurring IDR-macromolecule or an IDR-polypeptide according to any one of claims 41 to 51.

57. A kit according to claim 56, further comprising additional RPA components comprising an RPA recombinase agent, and / or an RPA recombinase loading protein, 10 and / or polymerase, and / or first and second nucleic acid primers, and / or an exonuclease, and / or a buffer, and / or a source of multivalent metal ions, preferably divalent metal cations.

58. A kit according to claim 56 or claim 57, wherein all components are provided in 15   lyophilized form.

59. A process of stimulating or enhancing liquid-liquid demixing in a solution, the process comprising providing a solution comprising an IDR-macromolecule or an IDR-polypeptide according to any one of claims 41 to 51, and contacting the IDR-20 macromolecule or IDR-polypeptide in solution with multivalent metal ions whereupon liquid-liquid demixing in the solution is stimulated or enhanced.

60. A process according to claim 59, wherein liquid-liquid demixing results in the formation of detectable phase-separated particles, in the solution.2561. A process according to claim 59 or claim 60, wherein the multivalent metal ions are divalent metal ions, optionally Mg2+, Mn2+, Ca2+, Co2+ or Ni2+, preferably Mg2+, Mn2+ or Ca2+, more preferably Mg2+.2026234101   18 Sep 202662. The use of multivalent metal ions in stimulating or enhancing liquid-liquid demixing in a solution, said demixing being mediated by an IDR-macromolecule or an IDR-polypeptide according to any one of claims 41 to 51.5   63. The use according to claim 62, wherein liquid-liquid demixing results in theformation of detectable phase-separated particles in the solution.

64. The use according to claim 62 or claim 63, wherein the multivalent metal ions are divalent metal ions, optionally Mg2+, Mn2+, Ca2+, Co2+ or Ni2+, preferably Mg2+, 10   Mn2+ or Ca2+, more preferably Mg2+.

65. The use according to any one of claims 62 to 64, wherein the multivalent metal ions engage in aromatic cation-pi interactions with amino acid residues in the one or more functional IDR amino acid sequences, thereby promoting liquid-liquid demixing.1566. A non-naturally occurring IDR-macromolecule or an IDR-polypeptide according to any one of claims 41 to 48 for use in therapy or for use as a diagnostic agent.

67. A method for determining the nucleotide sequence of one or more target20 polynucleotide molecules, said method comprising the steps of:(i) performing a process according to any one of claims 1 to 40 to amplify the one or more target polynucleotide molecules thereby obtaining a population comprising a plurality of copies of the one or more target polynucleotide molecules; and(ii) performing one or more nucleic acid sequencing reactions on the 25 population comprising the plurality of copies of the target polynucleotide molecules, preferably wherein the method is performed in a solid phase reaction system which comprises a surface.

68. The use of an IDR-macromolecule or an IDR-polypeptide according to any one 30 of claims 41 to 51 in a method for determining the nucleotide sequence of one or more2026234101   18 Sep 2026target polynucleotide molecules, preferably DNA molecules, preferably wherein the method is as defined in claim 67.

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