CN111801428A - 一种获得单细胞mRNA序列的方法 - Google Patents

一种获得单细胞mRNA序列的方法 Download PDF

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CN111801428A
CN111801428A CN201880083669.8A CN201880083669A CN111801428A CN 111801428 A CN111801428 A CN 111801428A CN 201880083669 A CN201880083669 A CN 201880083669A CN 111801428 A CN111801428 A CN 111801428A
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CN111801428B (zh
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范飞
程小芳
章文蔚
汪为茂
崔路漫
王欧
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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
    • C12Q1/6869—Methods for sequencing
    • 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
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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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
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6816—Hybridisation assays characterised by the detection means
    • C12Q1/6818—Hybridisation assays characterised by the detection means involving interaction of two or more labels, e.g. resonant energy transfer
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    • 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
    • C12Q2565/00—Nucleic acid analysis characterised by mode or means of detection
    • C12Q2565/50—Detection characterised by immobilisation to a surface
    • C12Q2565/514—Detection characterised by immobilisation to a surface characterised by the use of the arrayed oligonucleotides as identifier tags, e.g. universal addressable array, anti-tag or tag complement array

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Abstract

本发明公开了一种获得单细胞mRNA序列的方法。本发明的方法：(1)利用细胞标签载体捕获细胞的mRNA，反转录得到具有细胞标签的cDNA；来自于同一个细胞的cDNA具有相同的细胞标签，来自于不同细胞的cDNA具有不同的细胞标签；(2)利用转座酶复合体和分子标签载体，得到具有分子标签的多个cDNA片段；来自于同一cDNA的各个片段具有相同的分子标签，来自于不同cDNA的片段具有不同的分子标签；(3)高通量测序；(4)根据分子标签进行序列拼接，得到每个mRNA的序列；根据细胞标签，得到每个单细胞的所有mRNA的序列。本发明提供的方法可用于高通量获得大量单细胞中每个单细胞的所有mRNA的序列。

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PCT国内申请，说明书已公开。

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  1. PCT国内申请，权利要求书已公开。
CN201880083669.8A 2017-12-28 2018-12-27 一种获得单细胞mRNA序列的方法 Active CN111801428B (zh)

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CNPCT/CN2017/119342 2017-12-28
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PCT/CN2018/124293 WO2019129133A1 (zh) 2017-12-28 2018-12-27 一种获得单细胞mRNA序列的方法

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CN113106150A (zh) * 2021-05-12 2021-07-13 浙江大学 一种超高通量单细胞测序方法
CN115198000A (zh) * 2022-07-21 2022-10-18 北京寻因生物科技有限公司 一种单细胞全序列转录组文库构建的方法

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CN104032377A (zh) * 2014-06-30 2014-09-10 北京诺禾致源生物信息科技有限公司 单细胞转录组测序文库的构建方法及其应用
US20140272965A1 (en) * 2013-03-13 2014-09-18 Denovo Sciences, Inc. System and method for capturing and analyzing cells
US20160304952A1 (en) * 2015-04-14 2016-10-20 Massachusetts Institue Of Technology In situ nucleic acid sequencing of expanded biological samples
CN106754904A (zh) * 2016-12-21 2017-05-31 南京诺唯赞生物科技有限公司 一种cDNA的特异性分子标签及其应用
CN106795651A (zh) * 2014-06-26 2017-05-31 伊鲁米那股份有限公司 利用单管添加方案的加标签的核酸的文库制备
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

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EP3285062B1 (en) * 2012-02-29 2019-08-07 Fluidigm Corporation Methods, systems, and devices for multiple single-cell capturing and processing using microfluidics
US20160122753A1 (en) * 2013-06-12 2016-05-05 Tarjei Mikkelsen High-throughput rna-seq
US10975371B2 (en) * 2014-04-29 2021-04-13 Illumina, Inc. Nucleic acid sequence analysis from single cells
CN106498040B (zh) * 2016-10-12 2018-10-23 浙江大学 一种分子标记微珠及基于该分子标记微珠的高通量单细胞测序方法

Patent Citations (6)

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US20140272965A1 (en) * 2013-03-13 2014-09-18 Denovo Sciences, Inc. System and method for capturing and analyzing cells
CN106795651A (zh) * 2014-06-26 2017-05-31 伊鲁米那股份有限公司 利用单管添加方案的加标签的核酸的文库制备
CN104032377A (zh) * 2014-06-30 2014-09-10 北京诺禾致源生物信息科技有限公司 单细胞转录组测序文库的构建方法及其应用
US20160304952A1 (en) * 2015-04-14 2016-10-20 Massachusetts Institue Of Technology In situ nucleic acid sequencing of expanded biological samples
CN106754904A (zh) * 2016-12-21 2017-05-31 南京诺唯赞生物科技有限公司 一种cDNA的特异性分子标签及其应用
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

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113106150A (zh) * 2021-05-12 2021-07-13 浙江大学 一种超高通量单细胞测序方法
CN113106150B (zh) * 2021-05-12 2022-05-24 浙江大学 一种超高通量单细胞测序方法
CN114958996A (zh) * 2021-05-12 2022-08-30 浙江大学 一种超高通量单细胞测序试剂组合
CN115011679A (zh) * 2021-05-12 2022-09-06 浙江大学 一种超高通量单细胞测序方法
CN114958996B (zh) * 2021-05-12 2022-12-20 浙江大学 一种超高通量单细胞测序试剂组合
CN115198000A (zh) * 2022-07-21 2022-10-18 北京寻因生物科技有限公司 一种单细胞全序列转录组文库构建的方法

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EP3733865A4 (en) 2021-09-08
EP3733865A1 (en) 2020-11-04
CN111801428B (zh) 2023-05-23
WO2019129133A1 (zh) 2019-07-04
US20200318181A1 (en) 2020-10-08
US11718872B2 (en) 2023-08-08

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