CN105200041A - 用于构建单细胞转录组测序文库的试剂盒及文库构建方法 - Google Patents
用于构建单细胞转录组测序文库的试剂盒及文库构建方法 Download PDFInfo
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Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107475779A (zh) * | 2017-09-22 | 2017-12-15 | 上海美吉医学检验有限公司 | 适用于单细胞rrbs测序的文库建立方法及其应用 |
CN107488725A (zh) * | 2017-09-22 | 2017-12-19 | 上海美吉医学检验有限公司 | 适用于单细胞基因组甲基化测序的文库建立方法及其应用 |
CN108179174A (zh) * | 2018-01-15 | 2018-06-19 | 武汉爱基百客生物科技有限公司 | 一种高通量简化基因组测序文库的构建方法 |
CN108265047A (zh) * | 2016-12-30 | 2018-07-10 | 安诺优达基因科技(北京)有限公司 | 用于dna片段的非特异性复制的方法及试剂盒 |
CN109811045A (zh) * | 2017-11-22 | 2019-05-28 | 深圳华大智造科技有限公司 | 高通量的单细胞全长转录组测序文库的构建方法及其应用 |
CN109971843A (zh) * | 2017-12-27 | 2019-07-05 | 复旦大学泰州健康科学研究院 | 一种单细胞转录组的测序方法 |
WO2020135259A1 (zh) * | 2018-12-29 | 2020-07-02 | 阅尔基因技术(苏州)有限公司 | 一种测序文库构建试剂盒及其使用方法和应用 |
CN111378720A (zh) * | 2018-12-29 | 2020-07-07 | 浙江安诺优达生物科技有限公司 | 长链非编码rna的测序文库构建方法及其应用 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103667442A (zh) * | 2013-09-13 | 2014-03-26 | 西南民族大学 | 一种针对微量样本的转录组高通量测序方法 |
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 |
CN104894651A (zh) * | 2015-06-29 | 2015-09-09 | 天津诺禾医学检验所有限公司 | 微量起始dna的高通量测序文库构建方法及其所构建的高通量测序文库 |
-
2015
- 2015-10-22 CN CN201510696296.XA patent/CN105200041B/zh active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103667442A (zh) * | 2013-09-13 | 2014-03-26 | 西南民族大学 | 一种针对微量样本的转录组高通量测序方法 |
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 |
CN104894651A (zh) * | 2015-06-29 | 2015-09-09 | 天津诺禾医学检验所有限公司 | 微量起始dna的高通量测序文库构建方法及其所构建的高通量测序文库 |
Non-Patent Citations (2)
Title |
---|
CLONTECH LABORATORIES, INC.: "SMARTer® Universal Low Input RNA Kit for Sequencing UserManual", 《CLONTECH LABORATORIES, INC.》 * |
SIMONE PICELLI ET AL.: "Smart-seq2 for sensitive full-length transcriptome profiling in single cells", 《NATURE METHODS》 * |
Cited By (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108265047A (zh) * | 2016-12-30 | 2018-07-10 | 安诺优达基因科技(北京)有限公司 | 用于dna片段的非特异性复制的方法及试剂盒 |
CN108265047B (zh) * | 2016-12-30 | 2021-08-31 | 浙江安诺优达生物科技有限公司 | 用于dna片段的非特异性复制的方法及试剂盒 |
CN107475779A (zh) * | 2017-09-22 | 2017-12-15 | 上海美吉医学检验有限公司 | 适用于单细胞rrbs测序的文库建立方法及其应用 |
CN107488725A (zh) * | 2017-09-22 | 2017-12-19 | 上海美吉医学检验有限公司 | 适用于单细胞基因组甲基化测序的文库建立方法及其应用 |
CN109811045A (zh) * | 2017-11-22 | 2019-05-28 | 深圳华大智造科技有限公司 | 高通量的单细胞全长转录组测序文库的构建方法及其应用 |
CN109811045B (zh) * | 2017-11-22 | 2022-05-31 | 深圳华大智造科技股份有限公司 | 高通量的单细胞全长转录组测序文库的构建方法及其应用 |
CN109971843A (zh) * | 2017-12-27 | 2019-07-05 | 复旦大学泰州健康科学研究院 | 一种单细胞转录组的测序方法 |
CN108179174A (zh) * | 2018-01-15 | 2018-06-19 | 武汉爱基百客生物科技有限公司 | 一种高通量简化基因组测序文库的构建方法 |
WO2020135259A1 (zh) * | 2018-12-29 | 2020-07-02 | 阅尔基因技术(苏州)有限公司 | 一种测序文库构建试剂盒及其使用方法和应用 |
CN111378720A (zh) * | 2018-12-29 | 2020-07-07 | 浙江安诺优达生物科技有限公司 | 长链非编码rna的测序文库构建方法及其应用 |
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