CN109996892A - 单细胞测序文库的构建方法及其应用 - Google Patents

单细胞测序文库的构建方法及其应用 Download PDF

Info

Publication number
CN109996892A
CN109996892A CN201680091050.2A CN201680091050A CN109996892A CN 109996892 A CN109996892 A CN 109996892A CN 201680091050 A CN201680091050 A CN 201680091050A CN 109996892 A CN109996892 A CN 109996892A
Authority
CN
China
Prior art keywords
unicellular
fragmentation
chromatin
carried out
dna
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201680091050.2A
Other languages
English (en)
Other versions
CN109996892B (zh
Inventor
刘龙奇
刘传宇
吴亮
商周春
成梦南
袁月
徐礼钦
刘心
徐讯
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
MGI Tech Co Ltd
Original Assignee
MGI Tech Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by MGI Tech Co Ltd filed Critical MGI Tech Co Ltd
Publication of CN109996892A publication Critical patent/CN109996892A/zh
Application granted granted Critical
Publication of CN109996892B publication Critical patent/CN109996892B/zh
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • 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
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • 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
    • C12N15/1068—Template (nucleic acid) mediated chemical library synthesis, e.g. chemical and enzymatical DNA-templated organic molecule synthesis, libraries prepared by non ribosomal polypeptide synthesis [NRPS], DNA/RNA-polymerase mediated polypeptide synthesis
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • 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
    • C12N15/1096—Processes for the isolation, preparation or purification of DNA or RNA cDNA Synthesis; Subtracted cDNA library construction, e.g. RT, RT-PCR
    • 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
    • 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

Abstract

本发明提供了一种构建单细胞测序文库的方法，包括如下步骤:a)裂解单细胞，获得单细胞裂解物；b)对步骤a)所得单细胞裂解物进行细胞核与细胞质的分离，获得细胞核溶液与总RNA溶液；c)对步骤b)所得细胞核溶液进行染色质DNA文库构建，获得所述单细胞的染色质可接近性测序文库；并且，对步骤b)所得总RNA溶液进行转录组文库构建，获得所述单细胞的转录组测序文库。

Description

PCT国内申请，说明书已公开。

Claims (15)

  1. PCT国内申请，权利要求书已公开。
CN201680091050.2A 2016-12-07 2016-12-07 单细胞测序文库的构建方法及其应用 Active CN109996892B (zh)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2016/108940 WO2018103025A1 (zh) 2016-12-07 2016-12-07 单细胞测序文库的构建方法及其应用

Publications (2)

Publication Number Publication Date
CN109996892A true CN109996892A (zh) 2019-07-09
CN109996892B CN109996892B (zh) 2023-08-29

Family

ID=62490570

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201680091050.2A Active CN109996892B (zh) 2016-12-07 2016-12-07 单细胞测序文库的构建方法及其应用

Country Status (6)

Country Link
US (1) US11434483B2 (zh)
EP (1) EP3553180B1 (zh)
CN (1) CN109996892B (zh)
DK (1) DK3553180T3 (zh)
ES (1) ES2922281T3 (zh)
WO (1) WO2018103025A1 (zh)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110952147A (zh) * 2019-12-18 2020-04-03 南方科技大学 单细胞基因组测序用的dna文库的构建方法
CN113249439A (zh) * 2021-05-11 2021-08-13 杭州圣庭医疗科技有限公司 一种简化dna甲基化文库及转录组共测序文库的构建方法
CN115948621A (zh) * 2023-01-18 2023-04-11 珠海舒桐医疗科技有限公司 一种基于月经血dna的hpv筛查方法

Families Citing this family (39)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9701998B2 (en) 2012-12-14 2017-07-11 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10752949B2 (en) 2012-08-14 2020-08-25 10X Genomics, Inc. Methods and systems for processing polynucleotides
US11591637B2 (en) 2012-08-14 2023-02-28 10X Genomics, Inc. Compositions and methods for sample processing
US20150376609A1 (en) 2014-06-26 2015-12-31 10X Genomics, Inc. Methods of Analyzing Nucleic Acids from Individual Cells or Cell Populations
US10323279B2 (en) 2012-08-14 2019-06-18 10X Genomics, Inc. Methods and systems for processing polynucleotides
US9951386B2 (en) 2014-06-26 2018-04-24 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10400280B2 (en) 2012-08-14 2019-09-03 10X Genomics, Inc. Methods and systems for processing polynucleotides
CA2894694C (en) 2012-12-14 2023-04-25 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10533221B2 (en) 2012-12-14 2020-01-14 10X Genomics, Inc. Methods and systems for processing polynucleotides
CA2900481A1 (en) 2013-02-08 2014-08-14 10X Genomics, Inc. Polynucleotide barcode generation
SG11201705615UA (en) 2015-01-12 2017-08-30 10X Genomics Inc Processes and systems for preparing nucleic acid sequencing libraries and libraries prepared using same
EP3262407B1 (en) 2015-02-24 2023-08-30 10X Genomics, Inc. Partition processing methods and systems
EP4029939B1 (en) 2017-01-30 2023-06-28 10X Genomics, Inc. Methods and systems for droplet-based single cell barcoding
US10400235B2 (en) 2017-05-26 2019-09-03 10X Genomics, Inc. Single cell analysis of transposase accessible chromatin
CN116064732A (zh) 2017-05-26 2023-05-05 10X基因组学有限公司 转座酶可接近性染色质的单细胞分析
WO2019084043A1 (en) 2017-10-26 2019-05-02 10X Genomics, Inc. METHODS AND SYSTEMS FOR NUCLEIC ACID PREPARATION AND CHROMATIN ANALYSIS
SG11201913654QA (en) 2017-11-15 2020-01-30 10X Genomics Inc Functionalized gel beads
US20200354767A1 (en) * 2017-11-16 2020-11-12 The Regents Of The University Of Colorado, A Body Corporate Methods to measure functional heterogeneity among single cells
US10829815B2 (en) 2017-11-17 2020-11-10 10X Genomics, Inc. Methods and systems for associating physical and genetic properties of biological particles
WO2019157529A1 (en) 2018-02-12 2019-08-15 10X Genomics, Inc. Methods characterizing multiple analytes from individual cells or cell populations
SG11202009889VA (en) 2018-04-06 2020-11-27 10X Genomics Inc Systems and methods for quality control in single cell processing
US11932899B2 (en) 2018-06-07 2024-03-19 10X Genomics, Inc. Methods and systems for characterizing nucleic acid molecules
WO2020009665A1 (en) * 2018-07-06 2020-01-09 Agency For Science, Technology And Research Method for single-cell transcriptome and accessible regions sequencing
CN109023537B (zh) * 2018-09-04 2021-10-08 上海交通大学 一种微量dna样品高通量测序文库的构建技术
EP3908673A1 (en) * 2019-01-07 2021-11-17 Agilent Technologies, Inc. Compositions and methods for genomic dna and gene expression analysis in single cells
US11845983B1 (en) 2019-01-09 2023-12-19 10X Genomics, Inc. Methods and systems for multiplexing of droplet based assays
EP3924505A1 (en) 2019-02-12 2021-12-22 10X Genomics, Inc. Methods for processing nucleic acid molecules
US11467153B2 (en) 2019-02-12 2022-10-11 10X Genomics, Inc. Methods for processing nucleic acid molecules
JP7203276B2 (ja) * 2019-05-21 2023-01-12 深▲せん▼華大智造科技有限公司 メチル化されたｄｎａの標的領域に基づいてシーケンシングライブラリーを構築する方法及びキット
WO2021226917A1 (en) * 2020-05-14 2021-11-18 Singleron (Nanjing) Biotechnologies, Ltd. Novel method of one-step whole transcriptome amplification
CN111575348B (zh) * 2020-05-19 2024-01-09 广州微远医疗器械有限公司 宏基因组文库及建库方法和应用
CN112011596A (zh) * 2020-09-11 2020-12-01 深圳华大因源医药科技有限公司 一种同时检测多种微生物基因组的方法、溶液和试剂盒
CN114085897A (zh) * 2020-09-29 2022-02-25 生物岛实验室 空间组学测序方法
CN112280828A (zh) * 2020-10-22 2021-01-29 上海交通大学医学院 一种降低单细胞扩增偏倚性的离体组织细胞核分离方法
CN112941636A (zh) * 2021-04-02 2021-06-11 武汉博越致和生物科技有限公司 一种适用于简化转录组测序的文库构建方法及配套试剂盒
CN113462748A (zh) * 2021-05-11 2021-10-01 温氏食品集团股份有限公司 Dna测序文库的制备方法及试剂盒
CN113337590A (zh) * 2021-06-03 2021-09-03 深圳华大基因股份有限公司 一种二代测序方法和文库构建方法
CN114645074A (zh) * 2022-03-07 2022-06-21 华南农业大学 一种单花蕾微量rna建库方法
CN115612720A (zh) * 2022-09-28 2023-01-17 上海欧易生物医学科技有限公司 一种适用于猕猴桃根组织单细胞测序的植物细胞核制备以及悬液优化方法

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104561244A (zh) * 2013-10-15 2015-04-29 天津大学 基于二代测序技术的微生物单细胞转录组分析方法
US20150291942A1 (en) * 2014-04-15 2015-10-15 Illumina, Inc. Modified transposases for improved insertion sequence bias and increased dna input tolerance
US20160060691A1 (en) * 2013-05-23 2016-03-03 The Board Of Trustees Of The Leland Stanford Junior University Transposition of Native Chromatin for Personal Epigenomics
CN105463089A (zh) * 2015-12-21 2016-04-06 同济大学 应用于斑马鱼胚胎的易接近转座酶核染色质高通量测序实验的方法

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS53122047A (en) 1977-03-30 1978-10-25 Sanden Corp Electromagnetic clutch
JPS5924538U (ja) 1982-08-09 1984-02-15 サンデン株式会社 電磁クラツチ
JP4174896B2 (ja) 1998-09-22 2008-11-05 株式会社デンソー 電磁クラッチ
KR20040034191A (ko) 2002-10-21 2004-04-28 한라공조주식회사 전자클러치용 디스크 및 허브 조립체
EP1486694B1 (en) 2003-06-10 2009-03-18 Halla Climate Control Corporation Disc and hub assembly for electromagnetic clutch in a compressor

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20160060691A1 (en) * 2013-05-23 2016-03-03 The Board Of Trustees Of The Leland Stanford Junior University Transposition of Native Chromatin for Personal Epigenomics
CN104561244A (zh) * 2013-10-15 2015-04-29 天津大学 基于二代测序技术的微生物单细胞转录组分析方法
US20150291942A1 (en) * 2014-04-15 2015-10-15 Illumina, Inc. Modified transposases for improved insertion sequence bias and increased dna input tolerance
CN105463089A (zh) * 2015-12-21 2016-04-06 同济大学 应用于斑马鱼胚胎的易接近转座酶核染色质高通量测序实验的方法

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
IAIN C MACAULAY等: "Separation and parallel sequencing of the genomes and transcriptomes of single cells using G&T-seq", 《NATURE PROTOCOLS》 *
JASON A REUTER等: "Simul-seq: combined DNA and RNA sequencing for whole-genome and transcriptome profiling", 《NAT METHODS》 *
SERENA LIU等: "Single-Cell transcriptome sequencing: recent advances and remaining challenges", 《F1000 FACULTY REV》 *
SIMONE PICELLI等: "Tn5 transposase and tagmentation procedures for massively scaled sequencing projects", 《GENOME RES》 *
WENFEI JIN等: "Genome-wide detection of DNase I hypersensitive sites in single cells and FFPE tissue samples", 《NATURE》 *
YU HOU等: "Single-cell triple omics sequencing reveals genetic, epigenetic and transcriptomic in hepatocellular carcinomas", 《CELL RESEARCH》 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110952147A (zh) * 2019-12-18 2020-04-03 南方科技大学 单细胞基因组测序用的dna文库的构建方法
CN110952147B (zh) * 2019-12-18 2023-05-05 南方科技大学 单细胞基因组测序用的dna文库的构建方法
CN113249439A (zh) * 2021-05-11 2021-08-13 杭州圣庭医疗科技有限公司 一种简化dna甲基化文库及转录组共测序文库的构建方法
CN115948621A (zh) * 2023-01-18 2023-04-11 珠海舒桐医疗科技有限公司 一种基于月经血dna的hpv筛查方法

Also Published As

Publication number Publication date
ES2922281T3 (es) 2022-09-12
US11434483B2 (en) 2022-09-06
EP3553180A4 (en) 2020-08-12
EP3553180A1 (en) 2019-10-16
US20190338279A1 (en) 2019-11-07
EP3553180B1 (en) 2022-05-04
WO2018103025A1 (zh) 2018-06-14
CN109996892B (zh) 2023-08-29
DK3553180T3 (da) 2022-07-25

Similar Documents

Publication Publication Date Title
CN109996892A (zh) 单细胞测序文库的构建方法及其应用
CN110050067B (zh) 产生经扩增的双链脱氧核糖核酸的方法以及用于所述方法的组合物和试剂盒
WO2020160044A1 (en) In-situ spatial transcriptomics
US8329887B2 (en) Synthesis of tagged nucleic acids
US11326201B2 (en) Method for removing non-target RNA from RNA sample
CN108138365A (zh) 一种高通量的单细胞转录组建库方法
US11634766B2 (en) Methods and compositions for analyzing nucleic acids
JP2010516284A (ja) マイクロｒｎａの検出のための方法、組成物及びキット
CN102971434A (zh) 一种甲基化dna的高通量测序方法及其应用
JP2022543051A (ja) 単一細胞分析
US20170233724A1 (en) Partitioning of DNA Sequencing Libraries into Host and Microbial Components
Bogdanova et al. Normalizing cDNA libraries
CN113025608A (zh) 细胞裂解液、试剂盒及应用
US20140272967A1 (en) Systems and methods for isolating nucleic acids
US11326160B2 (en) Method for making a cDNA library
CN111118126B (zh) 一种基于高通量测序的mRNA检测方法
CN108342385A (zh) 一种接头和通过高效率环化方式构建测序文库的方法
WO2023202030A1 (zh) 一种小分子rna的高通量测序文库构建方法
CN116287115A (zh) 一种获得单细胞或单细胞核样本的方法、使用其获得的样本及应用
WO2020135650A1 (zh) 一种基因测序文库的构建方法
WO2023159999A1 (zh) 一种单细胞开放染色质和转录组共测序文库的构建方法
CN117625595A (zh) 染色质可及性/转录组共检测试剂盒及其应用
CN107354148A (zh) 一种用于微量dna高效建库的方法
WO2024059622A2 (en) Methods for simultaneous amplification of dna and rna
WO2024028505A1 (en) Methods of preparing normalised nucleic acid samples, kits and devices for use in the method

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
CB02 Change of applicant information

Address after: 518083 the comprehensive building of Beishan industrial zone and 11 2 buildings in Yantian District, Shenzhen, Guangdong.

Applicant after: Shenzhen Huada Zhizao Technology Co.,Ltd.

Address before: 518083 the comprehensive building of Beishan industrial zone and 11 2 buildings in Yantian District, Shenzhen, Guangdong.

Applicant before: MGI TECH Co.,Ltd.

CB02 Change of applicant information
GR01 Patent grant
GR01 Patent grant