16 results about "Dna assembly" patented technology

Disclosed is a DNA assembly mix, comprising a 3′-5′ exonuclease enzyme which is XthA; and a buffer. Also disclosed is a DNA assembly mix, comprising a polymerase and ligase free composition comprising a 3′-5′ exonuclease enzyme; and a buffer. Also disclosed is a method of assembling a plurality of DNA fragments, comprising: (a) mixing the plurality of DNA fragments with the DNA assembly mix as disclosed herein; and (b) incubating the mixture from step (a) at a temperature for a period of time suitable for assembling the plurality of DNA fragments. Further disclosed is use of the DNA assembly mix as disclosed herein in high-throughput DNA assembly, wherein the DNA assembly mix is used in a microfluidic platform to assemble DNA.

The application provides a method for DNA sequencing by pigeon hash growth tree splicing and a computer readable medium. The application acquires a reads library, determines the length of a pigeon hash table to construct the pigeon hash table, and numbers the reads in the reads library; the head of each read and the tail of each read are obtained through row division fragment processing; each read of the reads library is stored in the pigeon hash table through iterative replacement storage; splicing growth is determined in combination with the pigeon hash table, information of the reads in the splicing growth process is stored in a dynamic pigeon hash table in combination with the position of the successfully matched read head, and DNA sequencing is realized through iterative execution of growth of the growth tree. The application expands when the size of the hash table is insufficient, thereby saving space resources. The application is suitable for DNA splicing sequencing engineering.

The application discloses a lock-key type DNA assembly for circulating tumor cell marker protein multiplex analysis, and belongs to the technical field of biomedical molecular diagnosis. The lock-key type DNA assembly comprises a lock type DNA assembly and a key type DNA. The lock type DNA assembly comprises a 16nt double-stranded displacement region, a 10nt single-stranded branch point region, a 7nt single-stranded closed region and a covalent double-stranded signal region containing a plurality of fluorescent groups. The key type DNA is a single-stranded sequence modified on an antibody. The application realizes in-situ signal amplification and CTC marker protein multiplex analysis on a single cell level based on the lock-key type DNA assembly design, provides a theoretical basis for circulating tumor cell multiplex detection analysis of cancer, and provides a new method for further realizing precise diagnosis and individualized treatment of cancer.

The present invention relates to recombinant DNA technology, in particular to methods for assembling two or more double stranded (ds) nucleic acid molecules with overlapping terminal sequences. In particular, the present invention relates to the use of a thermolabile DNA polymerase II derived 3′-5′ exonuclease isolated from Moritella viscoa and a thermolabile DNA polymerase I of marine origin in multi DNA assembly processes.

To provide an improved analysis method or the like capable of analysis using a specific bonding function of DNA.SOLUTION: The analysis method is an analysis method for analyzing a target substance in a sample using the sample that may contain the target substance, the analysis method including: Acquiring a first signal of the sample after the reaction, reacting the sample with a second test substance containing a DNA assembly consisting of DNA of one or more types of sequences and acquiring a second signal of the sample after the reaction, and analyzing the target substance in the sample based on a difference between the first signal and the second signal, wherein a dissociation constant between the DNA assembly and the target substance is larger than a dissociation constant between the aptamer and the target substance.SELECTED DRAWING: Figure 6

The invention discloses a key type DNA assembly for circulating tumor cell marker protein multiplex analysis, and belongs to the technical field of biomedical molecular diagnosis. The lock-key type DNA assembly comprises a lock type DNA assembly and key type DNA, the lock type DNA assembly comprises a 16nt double-strand displacement region, a 10nt single-strand fulcrum region, a 7nt single-strand closed region and a covalent double-strand signal region containing a plurality of fluorophores, and the key type DNA is a single-strand sequence modified on an antibody. According to the present invention, based on the key type DNA assembly design, the signal in-situ amplification and the multiple analysis of the CTC marker protein on the single cell level are achieved, the theoretical basis is provided for the multiple detection analysis of the cancer circulating tumor cells, and the new method is provided for the further realization of the accurate diagnosis and the personalized treatment of the cancer.

Provided is a method for the synthesis and assembly of large fragment genomic DNA in vitro. A double-stranded DNA sticky-end generation system is used in the method. The generation system comprises DNA assembly sequences, single-stranded guide DNA, nicking endonuclease A, nicking endonuclease B, and nicking endonuclease C. The DNA assembly sequence comprises, from the 5' end to the 3' end: a first nicking endonuclease recognition site unit, a target DNA sequence, and a second nicking endonuclease recognition site unit. The method comprises: dividing a genomic sequence into n target DNA sequences, preparing DNA assembly sequences, designing and synthesizing single-stranded guide DNA, and adding nicking endonuclease A, nicking endonuclease B, and nicking endonuclease C to each DNA assembly sequence and the single-stranded guide DNA for enzyme digestion; and performing in vitro DNA assembly on the enzyme-digested DNA assembly sequences. The method overcomes the limitation of short overhangs generated by restriction endonucleases and improves the success rate of large DNA molecule assembly.

The application belongs to the technical field of biotechnology, and particularly relates to a kind of multivalent lysosome targeting antibody platform based on DNA tetrahedron.The application constructs a kind of lysosome targeting chimera based on DNA tetrahedron, uses general-purpose secondary antibody as general-purpose binding site, is connected with commercial primary antibody, solves two major core defects of lack of aptamer resources in the prior art and the need to redesign DNA assembly sequence for new target, realizes the efficient degradation of different target proteins "plug and play";The chimera can still maintain structural integrity in complex physiological environment, and after connecting the commercial primary antibody, the target protein can be mediated by the lysosome pathway to realize non-consumption and continuous degradation, which greatly reduces the research and development cost and cycle of targeted protein degradation drugs, significantly improves the application flexibility and degradation efficiency, and is especially suitable for the preparation of antitumor drugs.

The invention relates to a nucleic acid for use in DNA assembly, wherein the nucleic acid comprises at least one methylation-protectable restriction element, the methylation-protectable restriction element comprising: a restriction enzyme recognition sequence that is recognised by a restriction enzyme that cleaves outside of the recognition sequence; and a DNA methylase recognition sequence, wherein the restriction enzyme recognition sequence and the DNA methylase recognition sequence overlap such that the base modified by the DNA methylase lies within the restriction enzyme recognition sequence, wherein the DNA methylase recognition sequence is not identical to or enclosed by the restriction enzyme recognition sequence, and wherein the DNA methylase recognition sequence does not overlap with the sequence that would form the overhang end sequence generated by the restriction enzyme. The invention further relates to asscociated methods and kits.

The application discloses a method for regulating catalytic behavior of 3'->5' exonuclease based on deoxyribose configuration adjustment and application thereof, and belongs to the technical field of nucleic acid chemistry, biological catalytic regulation and molecular engineering. In view of the problems of poor position accuracy and unpredictable effect of the existing nucleic acid enzyme regulation method, a hydrazine compound containing an NH-NH2 functional group is used to modify and reduce an AP site in DNA, so that the deoxyribose of the AP site is changed from a closed ring configuration to a stable open chain configuration. The DNA after the AP site is modified is incubated with a nucleic acid enzyme with 3'->5' exonuclease activity, so that the nucleic acid enzyme is specifically catalyzed to stop at a nucleotide downstream of the modified AP site, and the catalytic behavior of the nucleic acid enzyme is accurately and controllably regulated with single nucleotide resolution. The application does not need to rely on steric hindrance or enzyme molecular structure modification, the regulation effect is stable and predictable, and the application can be widely applied to high-resolution DNA damage positioning, controllable DNA assembly and DNA oxidative damage level detection.

The present invention allows for the isolation of sequence correct DNA fragments from heterogeneous mixtures of fragments through the implementation of several steps for the purification of DNA molecules, the amplification of said DNA molecules, identification of sequence correct DNA fragments, and isolation of fragments as templates for PCR-based production of further downstream fragments.

Provided herein are nanochannel devices fabricated in a substrate for holding an internal construct in a channel, wherein the construct comprises DNA, with the construct positioned so that a joinable end of the DNA is extending outside the channel into an accessible inlet reservoir. Also provided herein are methods for long DNA assembly, comprising introducing a shuttle with a block group in a nanochannel, introducing joinable DNA segment solution, performing a joining reaction, and advancing the shuttle in the channel while still leaving the tail in the channel.

The present invention discloses a DNA assembly molecular machine and its applications, relating to the field of molecular biotechnology. The DNA assembly molecular machine comprises a palm domain, two terminal insertion domains, a thumb domain, a finger domain, and an exonuclease activity domain. The DNA assembly molecule exhibits excellent stability and is suitable for isothermal amplification. Furthermore, the DNA assembly molecule and its mutants possess DNA assembly capabilities in the presence of various metal ions and at different temperatures, and can be applied to nanopore sequencing, single-molecule sequencing, RCA library construction, environmental testing, and in vitro diagnostics.

Provided herein are nanochannel devices fabricated in a substrate for holding an internal construct in a channel, wherein the construct comprises DNA, with the construct positioned so that a joinable end of the DNA is extending outside the channel into an accessible inlet reservoir. Also provided herein are methods for long DNA assembly, comprising introducing a shuttle with a block group in a nanochannel, introducing joinable DNA segment solution, performing a joining reaction, and advancing the shuttle in the channel while still leaving the tail in the channel.

The application discloses a one-step long-chain DNA parallel synthesis method based on nucleic acid framework space programming, and belongs to the fields of molecular biology and DNA nanotechnology. The method comprises the following steps: preparing a nucleic acid framework structure template with a specific template chain, wherein the nucleic acid framework structure is a two-dimensional or three-dimensional DNA assembly precisely programmed in space position; hybridizing the nucleic acid framework structure template with an oligonucleotide pool to form an assembly; and adding a nucleic acid ligase to sequentially connect the oligonucleotides, thereby obtaining a long-chain nucleic acid product. The application utilizes the nucleic acid framework structure to fix the precise position of the oligonucleotides, improves the local concentration and sequencing accuracy, and realizes one-step efficient connection. The application solves the problems of low fidelity, small flux and high cost in long-chain nucleic acid synthesis in the prior art, has the advantages of high synthesis efficiency, high fidelity, high-throughput parallel synthesis, strong scalability and the like, and is suitable for the fields of synthetic biology, genetic engineering and biological medicine research and development.

The invention provides annular long single-stranded ssDNA, a single-stranded DNA assembly and a preparation method and application of the annular long single-stranded ssDNA and the single-stranded DNA assembly. The annular long single-stranded ssDNA comprises an aptamer sequence and a DNA assembly skeleton sequence; wherein the aptamer sequence is used for identifying any biomacromolecules or micromolecules; basic groups in molecules of the annular long single-stranded ssDNA are complementarily paired and are self-assembled to form a single-stranded DNA assembly. The method comprises the following steps: preparing long single-stranded ssDNA (single-stranded deoxyribonucleic acid) of milligram to acrylic level by utilizing a biological fermentation method; due to the self-folding characteristic of the annular long single-stranded ssDNA, the aptamer functionalized single-stranded DNA nano assembly with highly accurate valence state and spatial distribution and excellent structural stability can be efficiently constructed. The obtained nano-scale single-stranded DNA assembly has a strong affinity effect on the new coronavirus, and can efficiently block the combination of the new coronavirus and host cells so as to inhibit virus infection. The preparation method has the advantages of low cost, stable product structure and excellent uniformity of products among batches, and is suitable for animal level new coronavirus infection inhibition application research.