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34 results about "T4 polynucleotide kinase" patented technology
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Kit and method for constructing DNA library
ActiveCN105603535AAchieve end repairAchieve plus ALibrary creationT4 polynucleotide kinasePolymerase L
The invention provides a kit and method for constructing a DNA library. The kit comprises an enzymic reagent, a buffer solution and a joint combination, wherein the enzymic reagent comprises a first enzyme mixed liquid, T4DNA ligase and a second enzyme mixed liquid, and the first enzyme mixed liquid is formed by mixing T4 Polynucleotide Kinase, T4DNA polymerase, a Klenow fragment and rTaq enzyme; the second enzyme mixed liquid is a 2X KAPA HiFi HotStart pre-mixed solution; the buffer solution comprises a first buffer solution and a second buffer solution, and the first buffer solution is formed by mixing a 10X T4DNA ligase buffer solution, dNTP mixed liquor and ATP; the second enzyme mixed liquid is formed by mixing ATP, Tris-HCL, MgCl2, PEG8000, Tween 20 and dithiothreitol; the joint combination comprises a joint sequence group and a tag sequence group. According to the kit, the library construction cost can be reduced and the amplification efficiency is improved.
Owner:BEIJING NOVOGENE TECH CO LTD
Sequencing library construction kit and using method and application thereof
ActiveCN109797436AImprove utilization efficiencyNucleotide librariesMicrobiological testing/measurementT4 polynucleotide kinaseOriginal data
The invention relates to the field of gene detection, in particular to a sequencing library construction kit and a using method and application thereof. The kit comprises restriction endonuclease, a buffer solution for a restriction endonuclease reaction, DNA ligase, a ligase buffer solution, a tail end repair reagent, a sequencing connector, a PCR primer and a PCR amplification system, wherein the tail end repair reagent comprises T4 polynucleotide kinase, T4 DNA polymerase, Klenow DNA polymerase, dNTPs and a repair buffer solution. The sequence of a genome obtained from the kit is largely derived from different positions (intervals of 500 bp or above) on an original genome, so that the utilization efficiency of original data can be improved by 150% or above when the kit is used for analyzing the copy number variation of the genome.
Owner:CARRIER GENE TECH SUZHOU CO LTD
Silver nanocluster fluorescent probe based T4 polynucleotide kinase activity detecting method
InactiveCN108823280AWide detection rangeLow detection limitMicrobiological testing/measurementFluorescence/phosphorescenceT4 polynucleotide kinaseNucleotide kinase activity
Owner:THE SECOND HOSPITAL OF NANJING
Novel method for beacon-free detection of T4 PNKP (T4 polynucleotide kinase)/phosphatase and inhibitor of T4 PNKP/phosphatase on basis of fluorescent copper nanoparticles
ActiveCN104894222ALow costSimple and fast operationMicrobiological testing/measurementT4 polynucleotide kinaseSodium ascorbate
The invention discloses a novel method for beacon-free detection of T4 PNKP (T4 polynucleotide kinase) / phosphatase and an inhibitor of T4 PNKP / phosphatase on the basis of fluorescent copper nanoparticles. The method is implemented according to steps as follows: 1), a hairpin probe modified by 3'-terminal phosphorylation is designed, and 5' terminal is a sequence containing a base T; 2), the probe is diluted to the concentration being 100-1,000 nM with a Tris-HCl buffer solution, certain quantities of T4 PNKP, Klenow Fragment and dNTPs are mixed with the probe, and the mixture reacts at the temperature of 37 DEG C for 40-150 minutes; 3), the reaction liquid is diluted to form an MOPS buffer system, and copper sulfate and sodium ascorbate are added and react for 1-30 minutes at the room temperature; 4), the fluorescence strength of the finally obtained reaction liquid is detected with a fluorescence instrument, and a corresponding relation between the fluorescence strength of the system and T4 PNKP concentration is obtained. According to the method, the operation is simple, the cost is low, the sensitivity is high, the selectivity is good, T4 PNKP and the inhibitor can be quantitatively detected and screened, and a potential application value is provided for establishment of other terminal modification enzyme detection methods and molecular biological diagnosis.
Owner:郑州亮点生物技术有限公司
Method for quantitatively determining activity of T4 polynucleotide kinase
InactiveCN104293929AEasy to operateGood repeatabilityMicrobiological testing/measurementT4 polynucleotide kinaseRadioactive contamination
The invention discloses a method for quantitatively determining activity of T4 polynucleotide kinase. The method for quantitatively determining the activity of T4 polynucleotide kinase comprises the following steps: in a reaction system used for phosphorylation, adding ATP by taking oligonucleotide as a substrate for phosphorylating an end 5' of oligonucleotide by utilizing T4 polynucleotide kinase, so as to generate phosphorylated oligonucleotide; then adding a luciferase reporter gene system into the reaction system, wherein luciferase in the luciferase reporter gene system utilizes residual ATP in the reaction system to oxidize luciferin to form oxygenated luciferase and produce chemiluminiscence at the same time; and detecting numerical value of luminescence, and deriving activity degree of T4 polynucleotide kinase by virtue of detection results, wherein the activity degree of T4 polynucleotide kinase is in negative correlation with luminous quantity. Compared with the prior art, the method for quantitatively determining the activity of T4 polynucleotide kinase has the advantages that no radioactive contamination is produced, operation is easy, and quantitative detection analysis can be carried out on the activity of T4 polynucleotide kinase.
Owner:VAZYME BIOTECH NANJING
Method for performing DNA terminal repair/dA addition by adopting one-step method and application
InactiveCN105567684AAvoid lostReduce consumptionNucleotide librariesDNA/RNA fragmentationT4 polynucleotide kinaseEnzyme system
The invention belongs to the technical field of molecular biology and particularly relates to a method for performing DNA terminal repair / dA addition by adopting a one-step method and application thereof. The method adopts an enzyme system of T4DNA polymerase, Taq DNA polymerase and T4 polynucleotide kinase. The time required by the DNA terminal repair / dA addition in the method is shortened from one and a half hours to 40 minutes, and the reagent consumable consumption is reduced due to the fact that a DNA purification step is omitted, the cost is saved, DNA loss caused by purification is avoided, the preparation efficiency of a DNA library is improved, and the lowest DNA quality required by preparation of the DNA library is reduced.
Owner:SHANGHAI PASSION BIOTECHNOLOGY CO LTD
Method for determining activity of T4 polynucleotide kinase based on magnetic nano material and biological signal amplification technology
ActiveCN112710710AHigh sensitivityHigh selectivityMicrobiological testing/measurementMaterial analysis by electric/magnetic meansElectrochemical responseT4 polynucleotide kinase
The invention discloses a method for determining the activity of T4 polynucleotide kinase based on a magnetic nano material and a biological signal amplification technology. Fe3O4@TiO2 magnetic core-shell nanoparticles are prepared by adopting a hydrothermal method; when ATP and T4 PNK exist, the 5'end of a rolling circle amplification reaction primer chain S1 is phosphorylated and modified to the surfaces of the Fe3O4@TiO2 magnetic nanoparticles; an annular template S2 is added, and a nucleic acid chain S2 is hybridized with the Fe3O4@TiO2 magnetic nanoparticle surface primer chain S1 to form an annular mixture; an RCA reaction is carried out on the surfaces of the Fe3O4@ TiO2 magnetic nanoparticles; a ferrocene-labeled nucleic acid chain S3 is added, the Fc-S3 is hybridized with an RCA reaction product, an obtained product is modified onto the surfaces of the magnetic nanoparticles, and an electrochemical response signal is enhanced; and through magnetic enrichment of the GME and the electrochemical response signal, high-sensitivity and quantitative determination of the activity of T4 PNK is realized.
Owner:YUNNAN MINZU UNIV
Non-invasive prenatal detection kit for down syndrome
The invention provides a non-invasive prenatal detection kit for down syndrome, which comprises a T4 DNA ligase buffer solution, a Klenow fragment (5 units / mul), T4 polynucleotide kinase (T4PNK), an linker sequence, a DNA (deoxyribonucleic acid) sequencing primer, a PCR (polymerase chain reaction) component, a DNA sequencing component and the like. Through establishing an in vitro DNA library from peripheral blood of a pregnant woman, the kit can be used for accurate sequencing a genomic sequence of a fetus and screening the down syndrome of the fetus, with accurate results, and without false positive and false negative. Furthermore, the kit can be used for detection of a small number of in vitro peripheral blood samples, so as to greatly reduce the pain and risk of the pregnant woman, and the kit has good application prospects.
Owner:解码(上海)生物医药科技有限公司
Quantitative detection method of T4 polynucleotide kinase
ActiveCN110487867AHigh sensitivityGood choiceMaterial analysis by electric/magnetic meansT4 polynucleotide kinasePhosphate
The invention discloses a quantitative detection method of T4 polynucleotide kinase. The quantitative detection method comprises the steps of: designing a sulfhydrylated single-stranded DNA probe a, aprobe b and a probe c of which a 5'end is modified with amino, and fixing the probe a on the surface of a gold electrode through a gold-sulfhydryl bond, wherein the hydroxyl at the 5'end of the probea can be catalyzed and phosphorylated by T4 polynucleotide kinase, the DNA probe a is then hybridized with the probe b and the probe c, and the 3 'end of the probe c and a phosphate group at the 5' end of the probe a are subjected to a ligation reaction under the action of T4 DNA ligase; performing heating denaturating treatment, wherein the probe c is still retained on the surface of the electrode, the amino group at the tail end of the probe c can further capture the silver nanoparticles, and finally, the quantitative detection result of the T4 polynucleotide kinase is obtained by detectingthe stripping volt-ampere signal of the silver nanoparticles. The method has the characteristics of the high sensitivity, the good selectivity, the rapidness and the low cost, and the detection limitis 0.01 U / mL.
Owner:SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
Novel phosphorylated adenosine acylase as well as preparation method and application thereof
ActiveCN113512541ASolubleHigh expressionAntibody mimetics/scaffoldsTransferasesT4 polynucleotide kinaseNucleotide
The invention relates to a novel phosphorylated adenosine acylase and a preparation method and application thereof, prokaryotic inducible expression plasmid of Adenylase (PF0353) and T4 PNK (T4 Polynucleotide Kinase) fusion enzyme is constructed, the target fusion enzyme is obtained through expression and purification, the enzyme is high in water solubility and simple in expression and purification, the obtained fusion enzyme has the activity of T4 polynucleotide kinase and adenosine acylase at the same time, the 5'-hydroxy DNA can be subjected to the direct catalytic reaction in the presence of ATP to obtain the 5'-adenylate acylation DNA by using the one-step method, and the method for preparing the adenylate acylation DNA by using the fusion enzyme has advantages of low cost, high reaction efficiency, simpleness, convenience and rapidness, and can be flexibly used by the molecular biological technology.
Owner:WENZHOU MEDICAL UNIV
Recombinant T4 bacteriophage polynucleotide kinase (T4 PNK) and preparation method thereof
InactiveCN104178467AEasy to purifyEfficient purificationTransferasesFermentationSolubilityPurification methods
The invention relates to expression and purification of T4 bacteriophage polynucleotide kinase (T4 PNK) and particularly relates to soluble expression and a purification method of the T4 PNK, belonging to the field of biochemistry. According to the invention, a T4 PNK gene is cloned into a pGEX vector through vector construction, the T4 polynucleotide kinase and GST undergo fused expression under the condition of low temperature, then, the protein expression level reaches 15%, and the solubility reaches 95%.
Owner:孙启明
Construction method for short RNA fragment library, kit and application
ActiveCN112322700AMicrobiological testing/measurementLibrary creationT4 polynucleotide kinaseNucleotide
The invention relates to the field of gene sequencing, in particular to a construction method for a short RNA fragment library, a kit and application. The construction method comprises the following steps of carrying out pre-phosphorylation and dephosphorylation treatment on a short RNA fragment sample, so that a 3' terminal phosphate group in a short RNA fragment is modified into a hydroxyl group, and a 5' terminal hydroxyl group is modified into a phosphate group to obtain a first product; then connecting a 3' linker to obtain a first linker product; then carrying out dephosphorylation treatment to obtain a second product of which a 5' terminal is a phosphate group; then connecting a 5' linker to obtain a second linker product; and then carrying out reverse transcription and amplification to obtain the short RNA fragment library. The kit comprises T4 polynucleotide kinase and RNA 5' pyrophosphohydrolase. By utilizing the construction method and the kit, nucleic acid information of various short RNA fragments can be obtained; and the construction method and the kit are applied to the field of life science.
Owner:WUHAN BGI CLINICAL LAB CO LTD
Electrochemical analysis method for determining activity of T4 polynucleotide kinase based on magnetic nano material
ActiveCN112697858AHigh sensitivityEnhanced electrochemical signalMicrobiological testing/measurementMaterial analysis by electric/magnetic meansT4 polynucleotide kinaseMagnetite Nanoparticles
The invention discloses an electrochemical analysis method for determining the activity of T4 polynucleotide kinase based on a magnetic nano material, the method comprises the following steps: preparing Fe3O4@TiO2 magnetic core-shell nanoparticles by a hydrothermal method, phosphorylating the 5'end of a nucleic acid chain S1 when ATP and T4 PNK exist, and modifying the surface of the Fe3O4@TiO2 magnetic nanoparticles with the end phosphorylated S1 by specific reaction with TiO2; adding a gold nanoparticle-nucleic acid chain S2 compound (AuNPs-S2), and forming a Fe3O4@TiO2-S1 / S2-AuNPs compound due to a hybridization reaction between the nucleic acid chain S1 and the nucleic acid chain S2; using hexamino ruthenium [Ru(NH3)63+] is used as an electroactivity indicator, and realizing high-sensitivity, rapid and quantitative determination of the activity of T4PNK through magnetic enrichment and electrochemical signal enhancement on the surface of GME.
Owner:YUNNAN MINZU UNIV
Kit and method for constructing DNA library
PendingCN112708939AImprove the efficiency of database constructionNucleotide librariesLibrary creationT4 polynucleotide kinaseNucleotide
The invention provides a kit and a method for constructing a DNA library, the kit comprises a mixed enzyme solution, a mixed buffer solution, ligase and a ligation buffer solution, the mixed enzyme solution comprises endonuclease, T4 DNA polymerase, Klenow fragments, Taq DNA polymerase and T4 polynucleotide kinase, the mixed buffer solution and the ligation buffer solution comprise DTT and PEG 8000, the mixed enzyme solution and the mixed buffer solution are used for forming a first reaction system, the ligase and the ligation buffer solution are used for forming a second reaction system, in the first reaction system, the working concentration of the endonuclease is 0.01-0.05 U / mu L, the working concentration of the T4DNA polymerase is 0.04-0.1 U / mu L, the working concentration of the Klenow fragment is 0.005-0.02 U / mu L, the working concentration of the Taq DNA polymerase is 0.025-0.075 U / mu L, the working concentration of the T4 polynucleotide kinase is 0.1-0.5 U / mu L, the working concentration of the DTT is 5-10mM, and the working concentration of the PEG 8000 is 1-10%. According to the invention, the kit and the method for constructing the DNA library, which can improve the library construction efficiency, can be provided.
Owner:深圳海普洛斯医学检验实验室
Next-generation sequencing library building kit and method for improving library conversion rate
PendingCN112941635AImprove efficiencyReduce lossesMicrobiological testing/measurementLibrary creationT4 polynucleotide kinaseNucleotide
The invention relates to a method and a kit in the technical field of biology, and concretely relates to a next-generation sequencing library building kit and method for improving the library conversion rate. The kit and the method are particularly suitable for next-generation sequencing application of trace samples. The kit comprises: enzyme mixed solutions which comprises a first enzyme mixed solution, a second enzyme solution and a third enzyme mixed solution, wherein the first enzyme mixed solution is formed by mixing T4 polynucleotide kinase, T4 DNA polymerase, a Klenow fragment and Taq enzyme according to a ratio of 3: 1: 2: 1, the second enzyme solution is T4 DNA ligase, and the third enzyme mixed solution is a mixed solution of 2X QuarTaq HiFi HotStart; buffer solutions which comprise a first buffer solution corresponding to the first enzyme mixed solution and a second buffer solution corresponding to the second enzyme mixed solution, wherein the second buffer solution is formed by mixing Tris-HCl with the concentration of 0.1-0.6 M, MgCl2 with the concentration of 0.05-0.1 M, DTT with the concentration of 0.01-0.05 M, ATP with the concentration of 1-10 mM, PEG6000 with the concentration of 30%-50% and 1, 2 propylene glycol with the concentration of 10-20%; and a linker, an index sequence and a universal primer.
Owner:SHANGHAI DYNASTYGENE CO
cfDNA (circulating cell free DNA) terminal repairing enzyme composition, cfDNA terminal repairing buffer solution reagent, and construction method for sequencing library
ActiveCN111378632ALow end repair enzymesReduce dosageHydrolasesMicrobiological testing/measurementRibonucleosideT4 polynucleotide kinase
The invention provides a cfDNA (circulating cell free DNA) terminal repairing enzyme composition, which comprises the following enzymes, including an enzyme I and an enzyme II, wherein the enzyme I enables cfDNA to carry out DNA double-strand repairing and 3' terminal plus A when dNTP (deoxy-ribonucleoside triphosphate) is in the presence, and is a Taq DNA polymerase and Klenow Fragment, Exo-, andthe enzyme II enables a cfDNA 5' terminal to generate phosphorylation modification and is T4 Polynucleotide Kinase (T4 PNK for short). The invention also provides a cfDNA terminal repairing buffer solution reagent. The invention also provides a method which uses the above reagent to carry out cfDNA terminal repairing and construct a cfDNA sequencing library. The invention has the following technical effects that a use amount of reagent enzymes is small, cost is low, repairing efficiency is unexpectedly high, and therefore, the invention is suitable for carrying out terminal repairing on the cfDNA with a low sample amount.
Owner:JIANGSU COWIN BIOTECH CO LTD
Economical and practical nucleic acid chain 5'-hydroxyl phosphorylation method
The invention discloses an economical and practical nucleic acid chain 5'-hydroxyl phosphorylation method. The method comprises the following steps of taking purchased non-modified DNA (deoxyribonucleic acid) short fragments as raw materials, taking the 5'-end of the short fragments as hydroxyl, taking 10 times of a commercially purchased phosphorylation buffer solution component as a phosphoric acid source, and phosphorylating the 5'-hydroxyl of DNA under the action of T4 polynucleotide kinase. The method provided by the invention is wide in application range, mild in reaction condition, simple in post-treatment, high in recovery rate, low in cost and wide in application prospect.
Owner:PHARMARON BEIJING +1
Method for high-throughput rapid detection of pathogenic microorganisms
InactiveCN113215235AMicrobiological testing/measurementProteomicsGenomic sequencingT4 polynucleotide kinase
The invention discloses a method for detecting pathogenic microorganisms. The method comprises the following steps: simultaneously separating and extracting DNA and RNA from a sample of a subject; performing reverse transcription on the extracted RNA to obtain cDNA; treating the extracted DNA and the extracted cDNA by using a composition containing a plurality of enzymes, so as to obtain treated DNA and treated cDNA; performing PCR amplification on the basis of the treated DNA and the treated cDNA so as to construct a sequencing library; performing metagenome sequencing on the basis of the sequencing library to obtain a metagenome sequencing result of the sample; and determining the pathogenic microorganisms in the sample based on the metagenome sequencing result, a pathogenic microorganism genome database and a species classification database. The invention also discloses a kit for establishing the microbial metagenome sequencing library. The kit comprises a fragmentation and terminal repair system, wherein the fragmentation and terminal repair system comprises DNA incision enzymes, T4 polynucleotide kinase, T4 DNA polymerase, a Klinno fragment and Taq enzymes.
Owner:JIAXING YUNYING MEDICAL INSPECTION CO LTD
Cathode photoelectrochemical detection model of T4 polynucleotide kinase and application of cathode photoelectrochemical detection model
ActiveCN114199970AInhibition of immediate responseSimple and fast operationPreparing sample for investigationMaterial analysis by electric/magnetic meansT4 polynucleotide kinaseNucleotide
The invention relates to the field of analysis and detection, in particular to a detection model for non-labeled and non-fixed cathode photoelectrochemical detection of T4 polynucleotide kinase as well as a preparation method and application of the detection model. The construction method of the detection model comprises the following steps: (1) preparation of a Bi2O3 nano material; (2) preparation of a Bi2O3 / ITO electrode; (3) preparation of biological reaction solutions with different T4 polynucleotide kinase concentrations; (4) determination of light current; when the detection model is used for detecting the T4 PNK, biomolecule immobilization is not needed, the operation is simple and convenient, the cost is low, the selectivity is good, the sensitivity is high, the linear range is wide (5.0 * 10 <-4 >-10 U / mL), the detection limit is low (1.0 * 10 <-4 > U / mL), and the detection model has huge potential in practical application.
Owner:JIANGNAN UNIV
Method for building long-fragment target gene through polynucleotide kinase and application of method
InactiveCN110106196AEgobuyReduce financial burdenVector-based foreign material introductionT4 polynucleotide kinaseBiology
The invention belongs to the technical field of molecular biology, and particularly relates to a method for building a long-fragment target gene through polynucleotide kinase and application of the method. The method for building the long-fragment target gene through the polynucleotide kinase can effectively reduce the economic cost generated when the target gene is built and reduce the probability of base mutation. The method is characterized by comprising the steps that the target gene sequence is fragmented according to actual requirements, enzyme cutting site sequences are added to the twoends of each fragment respectively, and then corresponding target product sequence fragments are synthesized; by using the oligonucleotides 5' end of the T4 polynucleotide kinase, the target productis phosphorylated; the phosphorylated target product is annealed to obtain target fragments; a carrier vector is subjected to enzyme cutting, the formed sticky ends correspond to the enzyme cutting site sequences, an enzyme cutting product is recycled, and carrier fragments are obtained; the target fragments are connected with the carrier fragments to obtain the connection product.
Owner:WUHAN UNIV OF SCI & TECH
Methods and compositions for repair of DNA ends by multiple enzymatic activities
InactiveUS10301660B2Increase profitHigh sensitivityHydrolasesTransferasesT4 polynucleotide kinaseExonuclease III
Provided herein are compositions for and methods of generating ligation-competent nucleic acids. In some aspects, the compositions comprise Exonuclease III, T4 DNA Polymerase, Klenow, and / or T4 polynucleotide kinase.
Owner:TAKARA BIO USA INC
Enzyme composition, buffer reagent and sequencing library construction method for cfDNA end repair
ActiveCN111378632BLow end repair enzymesReduce dosageHydrolasesMicrobiological testing/measurementRibonucleosideT4 polynucleotide kinase
Owner:JIANGSU COWIN BIOTECH CO LTD
Method for determining activity of T4 polynucleotide kinase based on phosphate pillar [5] arene and carbon nanotube composite material
ActiveCN114839236AHigh selectivityHigh sensitivityMaterial electrochemical variablesT4 polynucleotide kinaseElectrochemical biosensor
The invention discloses a method for determining T4 polynucleotide kinase activity based on a phosphate pillar [5] arene and multiwalled carbon nanotube composite material. The method comprises the following steps: (1) designing a DNA oligonucleotide sequence; (2) synthesizing phosphate column [5] arene; (3) preparing a phosphate column [5] arene and multi-walled carbon nanotube composite material by adopting ultrasonic dispersion; (4) constructing an electrochemical biosensor; (5) determining the activity of T4 polynucleotide kinase; (6) determining a T4 polynucleotide kinase inhibitor; and (7) determining an actual sample. The method is simple to operate, low in cost, non-radioactive, high in sensitivity and good in selectivity, and has the characteristics of excellent host-guest identification characteristic based on the phosphate column [5] arene and large specific surface area, good stability and excellent conductivity of the multi-walled carbon nanotube.
![Method for determining activity of T4 polynucleotide kinase based on phosphate pillar [5] arene and carbon nanotube composite material](https://images-eureka.patsnap.com/patent_img/248bccf9-f12f-4f0c-a78d-ff5f0d0a50ea/HDA0003610161470000011.png "Method for determining activity of T4 polynucleotide kinase based on phosphate pillar [5] arene and carbon nanotube composite material")
![Method for determining activity of T4 polynucleotide kinase based on phosphate pillar [5] arene and carbon nanotube composite material](https://images-eureka.patsnap.com/patent_img/248bccf9-f12f-4f0c-a78d-ff5f0d0a50ea/HDA0003610161470000021.png "Method for determining activity of T4 polynucleotide kinase based on phosphate pillar [5] arene and carbon nanotube composite material")
![Method for determining activity of T4 polynucleotide kinase based on phosphate pillar [5] arene and carbon nanotube composite material](https://images-eureka.patsnap.com/patent_img/248bccf9-f12f-4f0c-a78d-ff5f0d0a50ea/HDA0003610161470000031.png "Method for determining activity of T4 polynucleotide kinase based on phosphate pillar [5] arene and carbon nanotube composite material")
Owner:YUNNAN MINZU UNIV
Method for measuring T4 polynucleotide kinase activity based on magnetic nanomaterials and biological signal amplification technology
ActiveCN112710710BHigh sensitivityHigh selectivityMicrobiological testing/measurementMaterial analysis by electric/magnetic meansElectrochemical responseT4 polynucleotide kinase
Owner:YUNNAN MINZU UNIV
Method for detecting polynucleotide kinase in high-salt high-protein biological sample
PendingCN114018890AReduce dosageReduce consumptionFluorescence/phosphorescenceAgainst vector-borne diseasesKinase activityT4 polynucleotide kinase
The invention discloses a novel method for detecting the activity of T4 polynucleotide kinase (T4 PNK) in complex biological matrix samples with high salt, high protein and the like by using silver nanoclusters (ssDNA / AgNCs) with stable oligonucleotide sequences. A large amount of salt and protein exist in a complex biological matrix sample, and the complex biological matrix sample needs to be subjected to high-multiple dilution, otherwise, fluorescence detection of T4 PNK is difficult to realize. Researches find that when enzymatic reactions such as T4 PNK and the like are completed, firstly, a quantitative AgNO3 solution is added, secondly, NaIO4 and Zn (NO3)2 solutions are added to oxidize protein, finally, dark-state ssDNA / AgNCs is added into the solution, bright-state ssDNA / AgNCs can be obtained after incubation, and fluorescence detection of the activity of T4 PNK is carried out. The method has the advantages of high detection sensitivity and high anti-interference capability, and can be suitable for high-salt and high-protein complex biological matrix samples without dilution.
Owner:TIANJIN NORMAL UNIVERSITY
Quantitative detection method of t4 polynucleotide kinase
ActiveCN110487867BHigh sensitivityGood choiceMaterial analysis by electric/magnetic meansT4 polynucleotide kinaseHybridization probe
The invention discloses a quantitative detection method for T4 polynucleotide kinase, which comprises the steps of: first designing a sulfhydrylated single-stranded DNA probe a, a probe b, and a probe c modified with an amino group at the 5' end, and the probe a is passed through The gold-sulfhydryl bond is immobilized on the surface of the gold electrode; the hydroxyl group at the 5' end of probe a can be catalyzed by T4 polynucleotide kinase to phosphorylate, DNA probe a then hybridizes with probe b and probe c, and the 3 of probe c The 'end and the phosphate group at the 5' end of probe a undergo a ligation reaction under the action of T4 DNA ligase; after heat denaturation treatment, probe c remains on the electrode surface, and the amino group at the end of probe c can further capture silver nanoparticles particles, and finally obtain the quantitative detection result of T4 polynucleotide kinase by detecting the stripping voltammetry signal of silver nanoparticles. The invention has the characteristics of high sensitivity, good selectivity, rapidity and low cost, and the detection limit is 0.01U / mL.
Owner:SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
A new method for label-free detection of t4 polynucleotide kinase/phosphatase and its inhibitors based on fluorescent copper nanoparticles
ActiveCN104894222BLow costSimple and fast operationMicrobiological testing/measurementT4 polynucleotide kinaseSodium ascorbate
The invention discloses a new method for detecting T4 polynucleotide kinase / phosphatase and its inhibitors based on fluorescent copper nanoparticles without labeling. Hairpin probe, the 5' end is a sequence rich in T bases; 2. Dilute the above probe with Tris-HCl buffer solution to a concentration of 100-1000nM, and add a certain amount of T4 PNKP, Klenow Fragment, and dNTPs to it Mix and react at 37°C for 40-150 minutes; 3. Dilute the above reaction solution into MOPS buffer system, add copper sulfate and sodium ascorbate, and react at room temperature for 1-30 minutes; 4. Use a fluorescence instrument to detect the fluorescence intensity of the final reaction solution. The corresponding relationship between the fluorescence intensity of the system and the concentration of T4 PNKP is obtained. The method of the present invention is simple in operation, low in cost, high in sensitivity and good in selectivity, and can quantitatively detect and screen T4 PNKP and its inhibitors. Establishment and molecular biology diagnostics offer potential applications.
Owner:郑州亮点生物技术有限公司
Nucleic acid molecule of codon-optimized T4 polynucleotide kinase and expression method of nucleic acid molecule
PendingCN114836448AGuaranteed outputGuaranteed purityBacteriaTransferasesT4 polynucleotide kinaseNucleotide
The invention discloses a nucleic acid molecule for coding T4 polynucleotide kinase, the nucleic acid molecule comprises a nucleotide sequence as shown in SEQ ID NO: 01, the invention further discloses a recombinant vector and a host cell containing the nucleotide sequence, an expression method of the recombinant vector and the host cell, and application of the produced T4 polynucleotide kinase. The nucleic acid molecule of the T4 polynucleotide kinase, disclosed by the invention, provides a technical guarantee for the yield, purity and activity of the T4 polynucleotide kinase.
Owner:ANNOROAD GENE TECH BEIJING +1
A novel phosphorylated adenylase and its preparation method and application
ActiveCN113512541BSolubleHigh expressionAntibody mimetics/scaffoldsTransferasesT4 polynucleotide kinaseNucleotide
A new type of phosphorylated adenylase and its preparation method and application, a prokaryotic inducible expression plasmid of Adenylase (PF0353) and T4 PNK (T4 Polynucleotide Kinase) fusion enzyme was constructed, and the target fusion enzyme was obtained through expression and purification, The enzyme has strong water solubility, simple expression and purification, and the obtained fusion enzyme has both the activities of T4 polynucleotide kinase and adenylase, and can directly catalyze the reaction of 5'-hydroxy DNA in the presence of ATP to obtain 5'-hydroxyl DNA in one step. 'Adenylylated DNA. The method for preparing adenylated DNA by using the fusion enzyme has low cost, high reaction efficiency, simplicity and rapidity, and can be used flexibly in molecular biology techniques.
Owner:WENZHOU MEDICAL UNIV
Enzyme composition for dna end repair, adenylation, phosphorylation
Enzyme compositions and their method of use that provide ready-to-use master mixtures. The compositions comprise a modified thermophilic DNA polymerase lacking 5′-3′ and 3′-5′ exonuclease activity premixed with T4 DNA polymerase, Klenow fragment and T4 polynucleotide kinase and all other necessary components, including reaction buffer and nucleoside triphosphates, required to perform DNA blunting, phosphorylation, and single nucleotide extension reactions in one tube and in two steps. Among other benefits, the mixture of different enzymes, buffers and nucleoside triphosphates is stable during prolonged storage.
Owner:THERMO FISHER SCI LITHUANIA HLDG UAB
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