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 PDFInfo
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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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- 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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- 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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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
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- 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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- 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。
Description
PCT国内申请，说明书已公开。
Claims (38)
- PCT国内申请，权利要求书已公开。
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2017119342 | 2017-12-28 | ||
| CNPCT/CN2017/119342 | 2017-12-28 | ||
| PCT/CN2018/124293 WO2019129133A1 (zh) | 2017-12-28 | 2018-12-27 | 一种获得单细胞mRNA序列的方法 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111801428A true CN111801428A (zh) | 2020-10-20 |
| CN111801428B CN111801428B (zh) | 2023-05-23 |
Family
ID=67066601
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201880083669.8A Active CN111801428B (zh) | 2017-12-28 | 2018-12-27 | 一种获得单细胞mRNA序列的方法 |
Country Status (4)
| Country | Link |
|---|---|
| US (1) | US11718872B2 (zh) |
| EP (1) | EP3733865B1 (zh) |
| CN (1) | CN111801428B (zh) |
| WO (1) | WO2019129133A1 (zh) |
Cited By (2)
| 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 |
| 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 |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114854827A (zh) * | 2022-05-18 | 2022-08-05 | 苏州极客基因科技有限公司 | 一种用于富集mRNA的磁珠复合物及其应用 |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| 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 |
| 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 |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3627140A1 (en) * | 2012-02-29 | 2020-03-25 | Fluidigm Corporation | Methods, systems, and devices for multiple single-cell capturing and processing using microfluidics |
| WO2014201273A1 (en) * | 2013-06-12 | 2014-12-18 | The Broad Institute, Inc. | 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 |
-
2018
- 2018-12-27 EP EP18896527.1A patent/EP3733865B1/en active Active
- 2018-12-27 WO PCT/CN2018/124293 patent/WO2019129133A1/zh not_active Ceased
- 2018-12-27 CN CN201880083669.8A patent/CN111801428B/zh active Active
-
2020
- 2020-06-23 US US16/909,366 patent/US11718872B2/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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)
| 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 |
Also Published As
| Publication number | Publication date |
|---|---|
| EP3733865A1 (en) | 2020-11-04 |
| WO2019129133A1 (zh) | 2019-07-04 |
| US11718872B2 (en) | 2023-08-08 |
| EP3733865B1 (en) | 2025-10-15 |
| EP3733865A4 (en) | 2021-09-08 |
| CN111801428B (zh) | 2023-05-23 |
| US20200318181A1 (en) | 2020-10-08 |
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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. |
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