WO2020258084A9 - 巢式多重pcr高通量测序文库制备方法及试剂盒 - Google Patents

巢式多重pcr高通量测序文库制备方法及试剂盒 Download PDF

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Publication number
WO2020258084A9
WO2020258084A9 PCT/CN2019/093066 CN2019093066W WO2020258084A9 WO 2020258084 A9 WO2020258084 A9 WO 2020258084A9 CN 2019093066 W CN2019093066 W CN 2019093066W WO 2020258084 A9 WO2020258084 A9 WO 2020258084A9
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primer
sequence
nested
multiplex pcr
throughput sequencing
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PCT/CN2019/093066
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French (fr)
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WO2020258084A1 (zh
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杨林
张艳艳
陈芳
蒋慧
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深圳华大智造科技有限公司
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Priority to CA3140403A priority Critical patent/CA3140403A1/en
Priority to EP19935242.8A priority patent/EP3992303A4/en
Priority to US17/617,854 priority patent/US20220235414A1/en
Priority to JP2021574288A priority patent/JP7281565B2/ja
Priority to CN201980095420.3A priority patent/CN113811620B/zh
Priority to AU2019453690A priority patent/AU2019453690B2/en
Priority to KR1020217042321A priority patent/KR20220011725A/ko
Priority to PCT/CN2019/093066 priority patent/WO2020258084A1/zh
Publication of WO2020258084A1 publication Critical patent/WO2020258084A1/zh
Publication of WO2020258084A9 publication Critical patent/WO2020258084A9/zh

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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
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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
    • C12Q1/6853—Nucleic acid amplification reactions using modified primers or templates
    • 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
    • C12Q1/6869—Methods for sequencing
    • C—CHEMISTRY; METALLURGY
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1065—Preparation or screening of tagged libraries, e.g. tagged microorganisms by STM-mutagenesis, tagged polynucleotides, gene tags
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    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/686—Polymerase chain reaction [PCR]
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    • 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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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
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  • Bioinformatics & Computational Biology (AREA)
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  • Plant Pathology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

一种巢式多重PCR高通量测序文库制备方法及试剂盒，巢式多重PCR高通量测序文库制备方法包括：使用正向引物和反向引物对目标区域进行扩增，正向引物是正向特异性序列，反向引物包括3'端的反向特异性序列和5'端的第一通用测序序列；对第一轮PCR扩增的产物进行纯化；使用巢式引物、第一标签引物和第二通用引物对纯化后的产物进行扩增，其中巢式引物位于正向引物的下游，巢式引物包括5'端的第二通用测序序列和3'端的特异性序列，第一标签引物3'端序列和反向引物5'端的部分或全部序列相同，且第一标签引物还包括第一标签序列，第二通用引物3'端序列和巢式引物5'端的部分或者全部序列相同。巢式多重PCR高通量测序文库制备方法能够有效避免多重PCR扩增过程中产生的二聚体，并且采用巢式扩增方法大大增加PCR扩增的特异性。
PCT/CN2019/093066 2019-06-26 2019-06-26 巢式多重pcr高通量测序文库制备方法及试剂盒 WO2020258084A1 (zh)

Priority Applications (8)

Application Number Priority Date Filing Date Title
CA3140403A CA3140403A1 (en) 2019-06-26 2019-06-26 Method for preparing nested multiplex pcr high-throughput sequencing library and kit
EP19935242.8A EP3992303A4 (en) 2019-06-26 2019-06-26 METHOD FOR BUILDING A HIGH-THROUGH-THROUGH NESTING MULTIPLEX PCR SEQUENCING LIBRARY AND KIT
US17/617,854 US20220235414A1 (en) 2019-06-26 2019-06-26 Method for preparing high-throughput sequencing library based on nested mutiplex pcr and kit for the same
JP2021574288A JP7281565B2 (ja) 2019-06-26 2019-06-26 ネストされたマルチプレックスｐｃｒハイスループットシーケンシングライブラリー調製方法及びキット
CN201980095420.3A CN113811620B (zh) 2019-06-26 2019-06-26 巢式多重pcr高通量测序文库制备方法及试剂盒
AU2019453690A AU2019453690B2 (en) 2019-06-26 2019-06-26 Method for preparing nested multiplex PCR high-throughput sequencing library and kit
KR1020217042321A KR20220011725A (ko) 2019-06-26 2019-06-26 네스티드 다중 pcr 고처리량 시퀀싱 라이브러리의 제조 방법 및 키트
PCT/CN2019/093066 WO2020258084A1 (zh) 2019-06-26 2019-06-26 巢式多重pcr高通量测序文库制备方法及试剂盒

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Application Number Priority Date Filing Date Title
PCT/CN2019/093066 WO2020258084A1 (zh) 2019-06-26 2019-06-26 巢式多重pcr高通量测序文库制备方法及试剂盒

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WO2020258084A1 WO2020258084A1 (zh) 2020-12-30
WO2020258084A9 true WO2020258084A9 (zh) 2021-02-04

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US (1) US20220235414A1 (zh)
EP (1) EP3992303A4 (zh)
JP (1) JP7281565B2 (zh)
KR (1) KR20220011725A (zh)
CN (1) CN113811620B (zh)
AU (1) AU2019453690B2 (zh)
CA (1) CA3140403A1 (zh)
WO (1) WO2020258084A1 (zh)

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CN114317696A (zh) * 2021-12-24 2022-04-12 深圳裕康医学检验实验室 一种试剂盒及其文库构建方法与污染检测方法
WO2024049276A1 (ko) * 2022-09-01 2024-03-07 주식회사 키오믹스 다중 표적 dna의 선택적 증폭용 조성물 및 이를 이용한 증폭 방법

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AU2019453690B2 (en) 2024-04-04
CN113811620B (zh) 2024-04-09
CA3140403A1 (en) 2020-12-30
JP2022537284A (ja) 2022-08-25
US20220235414A1 (en) 2022-07-28
KR20220011725A (ko) 2022-01-28
EP3992303A1 (en) 2022-05-04
WO2020258084A1 (zh) 2020-12-30
AU2019453690A1 (en) 2022-01-06
JP7281565B2 (ja) 2023-05-25
EP3992303A4 (en) 2023-03-15
CN113811620A (zh) 2021-12-17

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