CN107075561A - 一种核酸片段化方法和序列组合 - Google Patents

一种核酸片段化方法和序列组合 Download PDF

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CN107075561A
CN107075561A CN201480082162.2A CN201480082162A CN107075561A CN 107075561 A CN107075561 A CN 107075561A CN 201480082162 A CN201480082162 A CN 201480082162A CN 107075561 A CN107075561 A CN 107075561A
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sequence
nucleic acid
primer
extension
dosage
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韩鸿雁
耿春雨
郭冠瑛
章文蔚
蒋慧
江媛
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Shenzhen Hua Made Dazhi Technology Co Ltd
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BGI Shenzhen Co Ltd
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    • 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
    • C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
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    • 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
    • C12Q1/6853—Nucleic acid amplification reactions using modified primers or templates
    • C12Q1/6855—Ligating adaptors
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • 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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    • 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
    • C12Q2525/00—Reactions involving modified oligonucleotides, nucleic acids, or nucleotides
    • C12Q2525/10—Modifications characterised by
    • C12Q2525/179—Modifications characterised by incorporating arbitrary or random nucleotide sequences
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    • C12Q2525/00—Reactions involving modified oligonucleotides, nucleic acids, or nucleotides
    • C12Q2525/10—Modifications characterised by
    • C12Q2525/191—Modifications characterised by incorporating an adaptor
    • C—CHEMISTRY; METALLURGY
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    • C12Q2535/00—Reactions characterised by the assay type for determining the identity of a nucleotide base or a sequence of oligonucleotides
    • C12Q2535/122—Massive parallel sequencing

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  • Chemical & Material Sciences (AREA)
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  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

本发明公开了一种核酸片段化方法和序列组合，包括如下步骤：使用单链5’端延伸引物对变性后的核酸进行退火、延伸反应，其中单链5’端延伸引物包括5’端的测序平台接头序列和相连的随机序列，随机序列在变性后的核酸的随机位点发生退火；将双链3’端接头序列定向连接到延伸反应生成的核酸的3’端，变性纯化后得到片段化的包含两端接头序列的单链核酸。

Description

PCT国内申请，说明书已公开。

Claims (17)

  1. PCT国内申请，权利要求书已公开。
CN201480082162.2A 2014-10-13 2014-10-13 一种核酸片段化方法和序列组合 Pending CN107075561A (zh)

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PCT/CN2014/088509 WO2016058121A1 (zh) 2014-10-13 2014-10-13 一种核酸片段化方法和序列组合

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US (1) US10344317B2 (zh)
EP (1) EP3208343B1 (zh)
CN (1) CN107075561A (zh)
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Cited By (3)

* Cited by examiner, † Cited by third party
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WO2020191521A1 (zh) * 2019-03-22 2020-10-01 深圳华大智造科技有限公司 核酸序列、rna目标区域测序文库的构建方法及应用
CN113088561A (zh) * 2019-12-23 2021-07-09 深圳华大智造科技有限公司 构建侧翼已知序列的测序文库的引物组和方法
CN113088561B (zh) * 2019-12-23 2024-05-14 深圳华大智造科技股份有限公司 构建侧翼已知序列的测序文库的引物组和方法

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CN103930570A (zh) * 2011-09-16 2014-07-16 莱克斯奥根有限公司 核酸转录方法

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DE602004024034D1 (de) * 2003-01-29 2009-12-24 454 Corp Nukleinsäureamplifikation auf basis von kügelchenemulsion
WO2004081225A2 (en) * 2003-03-07 2004-09-23 Rubicon Genomics, Inc. Amplification and analysis of whole genome and whole transcriptome libraries generated by a dna polymerization process
JP2008546405A (ja) * 2005-06-23 2008-12-25 キージーン ナムローゼ フェンノートシャップ ハイスループットシーケンシング技術を使用して複雑なゲノムをシーケンシングするための改善された戦略
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020191521A1 (zh) * 2019-03-22 2020-10-01 深圳华大智造科技有限公司 核酸序列、rna目标区域测序文库的构建方法及应用
CN113088561A (zh) * 2019-12-23 2021-07-09 深圳华大智造科技有限公司 构建侧翼已知序列的测序文库的引物组和方法
CN113088561B (zh) * 2019-12-23 2024-05-14 深圳华大智造科技股份有限公司 构建侧翼已知序列的测序文库的引物组和方法

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EP3208343A4 (en) 2018-03-21
WO2016058121A1 (zh) 2016-04-21
EP3208343B1 (en) 2022-01-05
EP3208343A1 (en) 2017-08-23
US20180251813A1 (en) 2018-09-06
US10344317B2 (en) 2019-07-09

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