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1. CN111763664 - Enzyme reaction solution for constructing sequencing library and application of enzyme reaction solution

Office
China
Application Number 202010599034.2
Application Date 28.06.2020
Publication Number 111763664
Publication Date 13.10.2020
Grant Number 111763664
Grant Date 26.03.2021
Publication Kind B
IPC
C12N 9/16
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9Enzymes, e.g. ligases (6.); Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating, or purifying enzymes
14Hydrolases (3.)
16acting on ester bonds (3.1)
C12N 9/12
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9Enzymes, e.g. ligases (6.); Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating, or purifying enzymes
10Transferases (2.)
12transferring phosphorus containing groups, e.g. kinases (2.7)
C40B 50/06
CCHEMISTRY; METALLURGY
40COMBINATORIAL TECHNOLOGY
BCOMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES, IN SILICO LIBRARIES
50Methods of creating libraries, e.g. combinatorial synthesis
06Biochemical methods, e.g. using enzymes or whole viable microorganisms
CPC
C12N 9/16
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9Enzymes; Proenzymes; Compositions thereof
14Hydrolases (3)
16acting on ester bonds (3.1)
C12N 9/1252
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9Enzymes; Proenzymes; Compositions thereof
10Transferases (2.)
12transferring phosphorus containing groups, e.g. kinases (2.7)
1241Nucleotidyltransferases (2.7.7)
1252DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase
C12N 9/1276
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9Enzymes; Proenzymes; Compositions thereof
10Transferases (2.)
12transferring phosphorus containing groups, e.g. kinases (2.7)
1241Nucleotidyltransferases (2.7.7)
1276RNA-directed DNA polymerase (2.7.7.49), i.e. reverse transcriptase or telomerase
C12N 9/1241
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9Enzymes; Proenzymes; Compositions thereof
10Transferases (2.)
12transferring phosphorus containing groups, e.g. kinases (2.7)
1241Nucleotidyltransferases (2.7.7)
C40B 50/06
CCHEMISTRY; METALLURGY
40COMBINATORIAL TECHNOLOGY
BCOMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES, IN SILICO LIBRARIES
50Methods of creating libraries, e.g. combinatorial synthesis
06Biochemical methods, e.g. using enzymes or whole viable microorganisms
Applicants JIANGSU KANGKESI MEDICAL TECHNOLOGY CO., LTD.
江苏康科斯医疗科技有限公司
VAZYME BIOTECH CO., LTD.
南京诺唯赞生物科技股份有限公司
Inventors CAO LIN
曹林
NIE JUNWEI
聂俊伟
QU ZHIPENG
瞿志鹏
ZHANG LIJUN
张力军
HAN JINXIONG
韩锦雄
JIANG MINGYANG
江明扬
QIU CUI
邱翠
Agents 北京品源专利代理有限公司 11332
Title
(EN) Enzyme reaction solution for constructing sequencing library and application of enzyme reaction solution
(ZH) 一种用于构建测序文库的酶反应液及其应用
Abstract
(EN) The invention provides an enzyme reaction solution for constructing a sequencing library and an application of the enzyme reaction solution. The enzyme reaction solution comprises an enzyme composition and a reaction buffer solution, wherein the enzyme composition comprises endonuclease, DNA polymerase and polynucleotide kinase; and the reaction buffer solution comprises a metal salt, a substrateand a buffer medium aqueous solution. By optimizing the formula of the enzyme reaction solution and realizing cutting, tail end repairing and A tail adding treatment on a nucleic acid sample through aone-step reaction, in a proper buffer system, the enzyme digestion reaction rate and the tail end repairing reaction rate are balanced; and under the conditions that the initial amount of the sampleis 100 pg to 1 mu g and the treatment time is the same, a sequencing library with consistent length distribution is obtained.
(ZH) 本发明提供了一种用于构建测序文库的酶反应液及其应用,所述酶反应液包括酶组合物和反应缓冲液;所述酶组合物包括核酸内切酶、DNA聚合酶和多聚核苷酸激酶;所述反应缓冲液包括金属盐、底物和缓冲介质水溶液。本发明通过优化酶反应液的配方,通过一步反应实现对核酸样本的切割、末端修复和加A尾处理,在适宜的缓冲体系中,酶切反应速率和末端修复反应速率达到平衡,在样本起始量为100pg‑1μg、处理时间相同的情况下,获得了长度分布一致的测序文库。
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