CN115003867A - 一种待测样本rna的测序文库的构建方法 - Google Patents

一种待测样本rna的测序文库的构建方法 Download PDF

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Publication number
CN115003867A
CN115003867A CN202080094602.1A CN202080094602A CN115003867A CN 115003867 A CN115003867 A CN 115003867A CN 202080094602 A CN202080094602 A CN 202080094602A CN 115003867 A CN115003867 A CN 115003867A
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rna
dna fragment
library
primer
construction
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夏军
陈健
刘萍
张薇婷
杨林
史千玉
张理菁
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MGI Tech Co Ltd
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MGI Tech Co Ltd
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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
    • C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B50/00—Methods of creating libraries, e.g. combinatorial synthesis
    • C40B50/06—Biochemical methods, e.g. using enzymes or whole viable microorganisms

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Analytical Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Microbiology (AREA)
  • Wood Science & Technology (AREA)
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  • Physics & Mathematics (AREA)
  • Biotechnology (AREA)
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  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

本发明公开了一种待测样本RNA的测序文库的构建方法。该方法先使用Poly(A)聚合酶给RNA加上poly(A)，然后逆转录，之后使用末端转移酶给cDNA的3’端加上固定序列作为引物锚定位点，可以同时对多种RNA(包括mRNA、lncRNA、smallRNA、cfRNA和tRNA)进行建库，构建的文库中同时包括mRNA文库、lncRNA文库、smallRNA文库、cfRNA文库和tRNA文库，并且过程中不需要特殊的片段回收方法去回收smallRNA文库，使得对一份RNA样本只需要建一次库，就可以同时得到多种RNA的信息，从而提高的RNA建库效率和简便性。本发明具有重要的应用价值。

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PCT国内申请，说明书已公开。

Claims (14)

  1. PCT国内申请，权利要求书已公开。
CN202080094602.1A 2020-03-16 2020-03-16 一种待测样本rna的测序文库的构建方法 Pending CN115003867A (zh)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113355391A (zh) * 2021-06-04 2021-09-07 翌圣生物科技(上海)股份有限公司 一种用于靶向ffpe rna建库的方法

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WO2023046163A1 (zh) * 2021-09-26 2023-03-30 杭州诺辉健康科技有限公司 核酸文库构建和测序的方法和试剂盒
CN114457142A (zh) * 2022-02-25 2022-05-10 福建农林大学 一种高效加尾快速扩增植物dsRNA病毒基因组末端序列的方法及其应用
CN114736951A (zh) * 2022-04-20 2022-07-12 深圳大学 一种小分子rna的高通量测序文库构建方法
CN115976163A (zh) * 2022-09-14 2023-04-18 杭州联川生物技术股份有限公司 一种去除rRNA的探针组合物、建库试剂盒和建库方法
CN116200537B (zh) * 2022-10-14 2023-09-19 江苏省疾病预防控制中心(江苏省公共卫生研究院) 一种呼吸道合胞病毒a型全基因组测序方法
CN116042770B (zh) * 2022-11-01 2023-12-01 苏州京脉生物科技有限公司 尿液中miRNA文库制备和表达定量的方法及试剂盒

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Publication number Priority date Publication date Assignee Title
CN105463585B (zh) * 2014-09-12 2018-04-20 清华大学 基于单链dna分子构建测序文库的方法及其应用
CN107502607A (zh) * 2017-06-20 2017-12-22 浙江大学 一种大量组织、细胞样本mRNA的分子条形码标记、文库构建、测序的方法
CN109517881B (zh) * 2018-06-13 2021-07-13 清华大学 一种体液微量游离rna的高通量测序文库构建方法

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113355391A (zh) * 2021-06-04 2021-09-07 翌圣生物科技(上海)股份有限公司 一种用于靶向ffpe rna建库的方法

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