CN105714383B - 一种基于分子反向探针的测序文库构建方法和试剂 - Google Patents
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Classifications
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- 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
-
- 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
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- Chemical & Material Sciences (AREA)
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- Medicinal Chemistry (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Description
组分 | 用量 |
基因组DNA | 5-300ng |
MIP探针 | 1fmol |
Ampligase buffer | 2μL |
dNTP(10mM) | 2μL |
Ampligase | 1μL |
Phusion | 0.5μL |
水 | 补齐20μL |
组分 | 用量 |
Phusion MM | 25μL |
步骤2纯化后DNA | 10μL |
MIP Ad153_PCR2_1 | 5μL |
MIP Ad153_PCR2_3 | 5μL |
水 | 5μL |
组分 | 用量 |
DNA | 660ng |
MIP_Splint Oligo(10pmol/μL) | 10μL |
H2O | 补足70μL |
组分 | 用量 |
反应体系1 | 70μL |
ATP(0.1M) | 1.2μL |
10×TA Buffer | 1.2μL |
T4DNA Ligase | 1.2μL |
水 | 补足120μL |
组分 | 用量 |
反应体系2 | 120μL |
10×TA Buffer | 0.8μL |
Exo I | 3.9μL |
Exo III | 1.3μL |
Claims (12)
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
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CN2014108086477 | 2014-12-22 | ||
CN201410808647 | 2014-12-22 |
Publications (2)
Publication Number | Publication Date |
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CN105714383A CN105714383A (zh) | 2016-06-29 |
CN105714383B true CN105714383B (zh) | 2018-01-23 |
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Country Status (2)
Country | Link |
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CN (1) | CN105714383B (zh) |
HK (1) | HK1221710A1 (zh) |
Families Citing this family (25)
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CN106399519B (zh) * | 2016-09-30 | 2019-07-26 | 南京迪康金诺生物技术有限公司 | 一种基于发卡结构的目标区域捕获方法及其应用 |
WO2018214036A1 (zh) * | 2017-05-23 | 2018-11-29 | 深圳华大基因股份有限公司 | 基于滚环扩增的基因组目标区域富集方法及其应用 |
CN109593757B (zh) * | 2017-09-30 | 2021-08-03 | 厦门艾德生物医药科技股份有限公司 | 一种探针及其适用于高通量测序的对目标区域进行富集的方法 |
CN110021353B (zh) * | 2017-09-30 | 2020-11-06 | 厦门艾德生物医药科技股份有限公司 | 一种用于捕获富集基因组的特定区域的分子反向探针的筛选方法 |
CN109694908B (zh) * | 2017-10-24 | 2023-01-10 | 深圳乐土生物科技有限公司 | 检测耳聋基因20个突变位点的快速文库构建方法及检测方法 |
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JP6710427B2 (ja) * | 2017-12-29 | 2020-06-17 | アクト ゲノミクス (アイピー) カンパニー リミテッド | 配列アライメントおよびバリアントコールのための方法およびシステム |
CN110079592B (zh) * | 2018-01-26 | 2021-02-12 | 厦门艾德生物医药科技股份有限公司 | 用于检测基因突变和已知、未知基因融合类型的高通量测序靶向捕获目标区域的探针和方法 |
CN108396057B (zh) * | 2018-02-28 | 2021-11-09 | 重庆市肿瘤研究所 | 基于长链分子倒置探针的核酸靶向捕获测序文库制备方法 |
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CN114836415B (zh) * | 2022-05-06 | 2023-04-18 | 南京诺唯赞生物科技股份有限公司 | 一种mgi平台转座酶双端标签文库的制备 |
WO2023240611A1 (zh) * | 2022-06-17 | 2023-12-21 | 深圳华大智造科技股份有限公司 | 单链核酸环状文库的建库以及测序方法 |
CN115873927B (zh) * | 2022-08-04 | 2023-12-26 | 迈杰转化医学研究(苏州)有限公司 | 一种不依赖连接的核酸恒温滚环扩增方法及其应用 |
CN115873928B (zh) * | 2022-09-02 | 2024-01-09 | 迈杰转化医学研究(苏州)有限公司 | 一种基于双链环模板的滚环扩增方法及其应用 |
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- 2015-12-22 CN CN201510971452.9A patent/CN105714383B/zh active Active
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