Quartz-Seq2: a high-throughput single-cell RNA-sequencing method that effectively uses limited sequence reads

被引:100
作者
Sasagawa, Yohei [1 ]
Danno, Hiroki [1 ]
Takada, Hitomi [2 ]
Ebisawa, Masashi [1 ]
Tanaka, Kaori [1 ]
Hayashi, Tetsutaro [1 ]
Kurisaki, Akira [2 ]
Nikaido, Itoshi [1 ,3 ]
机构
[1] RIKEN, Adv Ctr Comp & Commun, Bioinformat Res Unit, Hirosawa 2-1, Wako, Saitama, Japan
[2] Nara Inst Sci & Technol, Grad Sch Biol Sci, Lab Stem Cell Technol, Takayama Cho 8916-5, Ikoma, Nara, Japan
[3] RIKEN, Ctr Dev Biol, Single Cell Omics Res Unit, Chuo Ku, 2-2-3 Minatojima Minamimachi, Kobe, Hyogo, Japan
基金
日本科学技术振兴机构;
关键词
High-throughput single-cell RNA-seq; Flow cytometry; Cell sorter; Quartz-Seq; Poly(A) tagging; Stromal vascular fraction; Mesenchymal stem cell; SEQ; PACKAGE; CYCLE;
D O I
10.1186/s13059-018-1407-3
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 [微生物学]; 090105 [作物生产系统与生态工程];
摘要
High-throughput single-cell RNA-seq methods assign limited unique molecular identifier (UMI) counts as gene expression values to single cells from shallow sequence reads and detect limited gene counts. We thus developed a high-throughput single-cell RNA-seq method, Quartz-Seq2, to overcome these issues. Our improvements in the reaction steps make it possible to effectively convert initial reads to UMI counts, at a rate of 30-50%, and detect more genes. To demonstrate the power of Quartz-Seq2, we analyzed approximately 10,000 transcriptomes from in vitro embryonic stem cells and an in vivo stromal vascular fraction with a limited number of reads.
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页数:24
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