CN108004301A - 基因目标区域富集方法及建库试剂盒 - Google Patents
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Description
DNA样本 | 60ul |
10×T4多核苷酸激酶缓冲液 | 10ul |
dNTP(每种10mM) | 2ul |
T4 DNA聚合酶 | 2ul |
T4多核苷酸激酶 | 2ul |
Klenow片段 | 2ul |
ddH2O | 22ul |
总体积 | 100ul |
连接纯化产物 | 40ul |
5×Q5Buffer | 20ul |
dNTP(10mM each) | 2ul |
Q5高保真DNA聚合酶(2U/μL) | 1ul |
探针1(10μM) | 1ul |
探针2(10μM) | 1ul |
ddH2O | 35ul |
总体积 | 100ul |
上一步纯化产物 | 30ul |
5×Q5 Buffer | 10ul |
dNTP(10mM each) | 1ul |
Q5高保真DNA聚合酶(2U/μL) | 1ul |
引物F(10μM) | 1.5ul |
引物R(10μM) | 1.5ul |
ddH2O | 5ul |
总体积 | 50ul |
总reads数 | 315060 |
比对到hg19上的reads数 | 296944 |
比对率 | 94.25% |
目标区域reads数 | 160349 |
目标区域reads比例 | 54% |
Claims (15)
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CN201711348235.XA CN108004301B (zh) | 2017-12-15 | 2017-12-15 | 基因目标区域富集方法及建库试剂盒 |
PCT/CN2018/080229 WO2019114146A1 (zh) | 2017-12-15 | 2018-03-23 | 基因目标区域富集方法及建库试剂盒 |
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CN201711348235.XA CN108004301B (zh) | 2017-12-15 | 2017-12-15 | 基因目标区域富集方法及建库试剂盒 |
Publications (2)
Publication Number | Publication Date |
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CN108004301A true CN108004301A (zh) | 2018-05-08 |
CN108004301B CN108004301B (zh) | 2022-02-22 |
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Country Status (2)
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CN (1) | CN108004301B (zh) |
WO (1) | WO2019114146A1 (zh) |
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CN108531582A (zh) * | 2018-05-15 | 2018-09-14 | 广州达瑞生殖技术有限公司 | 一种检测人类胚胎α-地中海贫血基因突变的引物组合及方法 |
CN109517819A (zh) * | 2018-10-24 | 2019-03-26 | 深圳市易基因科技有限公司 | 一种用于检测多靶点基因突变、甲基化修饰和/或羟甲基化修饰的检测探针、方法和试剂盒 |
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CN109825552A (zh) * | 2019-02-01 | 2019-05-31 | 厦门艾德生物医药科技股份有限公司 | 一种用于对目标区域进行富集的引物及方法 |
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CN110699426A (zh) * | 2019-01-02 | 2020-01-17 | 上海臻迪基因科技有限公司 | 基因目标区域富集方法及试剂盒 |
WO2020073748A1 (zh) * | 2018-10-11 | 2020-04-16 | 北京优乐复生科技有限责任公司 | 一种测序文库的构建方法 |
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CN112779320A (zh) * | 2020-12-04 | 2021-05-11 | 深圳市易基因科技有限公司 | 多区域dna甲基化检测探针设计及其检测方法 |
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WO2022156823A1 (zh) * | 2021-01-22 | 2022-07-28 | 上海羿鸣生物科技有限公司 | 一种优化的dna线性扩增方法及试剂盒 |
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