CN109689872A - 一种dna末端修复与加a的方法 - Google Patents

一种dna末端修复与加a的方法 Download PDF

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CN109689872A
CN109689872A CN201680088992.5A CN201680088992A CN109689872A CN 109689872 A CN109689872 A CN 109689872A CN 201680088992 A CN201680088992 A CN 201680088992A CN 109689872 A CN109689872 A CN 109689872A
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dna
polymerase
datp
reaction
connector
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CN109689872B (zh
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江媛
沈寒婕
李巧玲
魏汉敏
赵霞
张艳艳
王逸丛
陈芳
傅书锦
苏小珊
耿春雨
蒋慧
章文蔚
拉多杰·德马纳克
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MGI Tech Co Ltd
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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
    • C12N15/1093—General methods of preparing gene libraries, not provided for in other subgroups
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    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B50/00—Methods of creating libraries, e.g. combinatorial synthesis
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Abstract

一种DNA末端修复与加A的方法，该方法包括：在同一反应体系中，在dNTP存在下，利用聚合酶将片段化DNA的末端补平或切平，利用多聚核苷酸激酶将5’羟基转变成5’磷酸基团且将3’磷酸基团转变成3’羟基；在过量dATP存在下，利用不具有3’‑5’外切活性的聚合酶在双链DNA 3’末端加上dATP。所述方法能够在同一个反应体系中实现DNA末端修复与加A反应，简化酶反应和纯化步骤，提高起始DNA转化成可连接接头DNA的效率，降低对起始DNA总量的要求。

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PCT国内申请，说明书已公开。

Claims (21)

  1. PCT国内申请，权利要求书已公开。
CN201680088992.5A 2016-11-21 2016-11-21 一种dna末端修复与加a的方法 Active CN109689872B (zh)

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CN111394799A (zh) * 2020-03-11 2020-07-10 广州赛哲生物科技股份有限公司 一种脑膜炎病原体宏基因组二代测序文库的构建方法及其试剂盒
CN112176028A (zh) * 2019-07-05 2021-01-05 深圳华大生命科学研究院 一种基于核酸内切酶的快速wgs建库方法
CN113355379A (zh) * 2021-02-04 2021-09-07 康龙化成(北京)新药技术股份有限公司 一种经济实用的核酸链5’-羟基磷酸化方法

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CN109055518A (zh) * 2018-07-06 2018-12-21 天津华大医学检验所有限公司 用于常见遗传病检测的探针、基因芯片及制备方法和应用
CN110791813B (zh) * 2018-08-01 2023-06-16 广州华大基因医学检验所有限公司 对单链dna进行处理的方法及应用
CN109321567A (zh) * 2018-10-10 2019-02-12 菲鹏生物股份有限公司 测序用dna文库试剂盒以及测序用dna文库构建方法
US20220002713A1 (en) * 2018-10-11 2022-01-06 Beijing Euler Technology Limited Company Method for constructing sequencing library
CN109610008A (zh) * 2018-11-08 2019-04-12 广州华大基因医学检验所有限公司 基于高通量测序的中枢***感染病原检测文库构建方法、检测方法和试剂盒
CN113227370B (zh) * 2018-12-28 2024-04-09 江苏金斯瑞生物科技有限公司 一种单链dna合成方法
CN111073952B (zh) * 2018-12-29 2023-09-22 浙江安诺优达生物科技有限公司 构建dna文库的方法及其应用
CN109957605A (zh) * 2019-03-20 2019-07-02 嘉兴菲沙基因信息有限公司 适用于PacBio测序平台的海洋动物与软体动物的建库测序方法
CN109971827B (zh) * 2019-03-25 2020-05-01 纳昂达(南京)生物科技有限公司 血浆dna的建库方法和建库试剂盒
CN112301103B (zh) * 2019-08-02 2023-04-11 北京贝瑞和康生物技术有限公司 一种非特异性扩增天然短片段核酸的方法和试剂盒
CN112342627A (zh) * 2019-08-09 2021-02-09 深圳市真迈生物科技有限公司 一种核酸文库的制备方法及测序方法
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CN111139533B (zh) * 2019-09-27 2021-11-09 上海英基生物科技有限公司 稳定性增加的测序文库接头
CN110656156A (zh) * 2019-10-14 2020-01-07 湖南大地同年生物科技有限公司 一种超低频突变核酸片段检测方法、文库构建方法、引物设计方法和试剂
CN110846382B (zh) * 2019-11-29 2023-02-28 北京科迅生物技术有限公司 胎儿游离dna的富集方法
CN110846724B (zh) * 2019-12-03 2023-04-28 江西海普洛斯医学检验实验室有限公司 构建mRNA链特异文库的方法及试剂盒
CN111378632B (zh) * 2020-03-19 2021-03-19 江苏康为世纪生物科技股份有限公司 cfDNA末端修复的酶组合物、缓冲液试剂和测序文库的构建方法
CN113774120B (zh) * 2020-06-09 2024-05-14 深圳华大智造科技股份有限公司 Dnb双末端测序方法
EP4170028A4 (en) * 2020-06-19 2023-10-18 MGI Tech Co., Ltd. HIGH COMPATIBILITY PCR-FREE LIBRARY CONSTRUCTION AND SEQUENCING METHOD
CN111763664B (zh) * 2020-06-28 2021-03-26 江苏康科斯医疗科技有限公司 一种用于构建测序文库的酶反应液及其应用
CN112226821B (zh) * 2020-10-16 2024-02-06 鲲羽生物科技(江门)有限公司 一种基于双链环化的mgi测序平台测序文库的构建方法
CN112708619B (zh) * 2020-12-30 2022-05-17 纳昂达(南京)生物科技有限公司 Mgi平台的建库用接头、试剂盒及建库方法
CN112708939A (zh) * 2020-12-30 2021-04-27 深圳海普洛斯医学检验实验室 构建dna文库的试剂盒及方法
CN112779315A (zh) * 2021-01-04 2021-05-11 深圳海普洛斯医学检验实验室 一种基于Illumina测序平台的游离DNA文库构建用试剂盒及其应用
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CN112176028A (zh) * 2019-07-05 2021-01-05 深圳华大生命科学研究院 一种基于核酸内切酶的快速wgs建库方法
CN112176028B (zh) * 2019-07-05 2024-04-16 深圳华大生命科学研究院 一种基于核酸内切酶的快速wgs建库方法
CN111394799A (zh) * 2020-03-11 2020-07-10 广州赛哲生物科技股份有限公司 一种脑膜炎病原体宏基因组二代测序文库的构建方法及其试剂盒
CN113355379A (zh) * 2021-02-04 2021-09-07 康龙化成(北京)新药技术股份有限公司 一种经济实用的核酸链5’-羟基磷酸化方法

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