Quantitative assessment of single-cell RNRNA-sequencing methods

被引：520

作者：

Wu, Angela R. ^{[1
]}

Neff, Norma F. ^{[1
]}

Kalisky, Tomer ^{[1
]}

Dalerba, Piero ^{[2
,3
,4
]}

Treutlein, Barbara ^{[1
]}

Rothenberg, Michael E. ^{[5
]}

Mburu, Francis M. ^{[1
,6
]}

Mantalas, Gary L. ^{[1
]}

Sim, Sopheak

Clarke, Michael F. ^{[2
,3
,4
]}

Quake, Stephen R. ^{[1
,6
,7
]}

机构：

[1] Stanford Univ, Dept Bioengn, Stanford, CA 94305 USA

[2] Stanford Univ, Ctr Med, Dept Med, Div Oncol, Stanford, CA 94305 USA

[3] Stanford Univ, Ctr Med, Inst Stem Cell Biol & Regenerat Med, Stanford, CA 94305 USA

[4] Stanford Univ, Ctr Med, Ludwig Canc Ctr, Stanford, CA 94305 USA

[5] Stanford Univ, Ctr Med, Dept Med, Div Gastroenterol & Hepatol, Stanford, CA 94305 USA

[6] Howard Hughes Med Inst, Stanford, CA USA

[7] Stanford Univ, Dept Appl Phys, Stanford, CA 94305 USA

来源：

NATURE METHODS | 2014年 / 11卷 / 01期

基金：

美国国家卫生研究院;

关键词：

MESSENGER-RNA-SEQ; SPLICE JUNCTIONS; STEM-CELLS; EXPRESSION; AMPLIFICATION; LANDSCAPE; ALIGNMENT;

D O I：

10.1038/nmeth.2694

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Interest in single-cell whole-transcriptome analysis is growing rapidly, especially for profiling rare or heterogeneous populations of cells. We compared commercially available single-cell RNRNA amplification methods with both microliter and nanoliter volumes, using sequence from bulk total RNRNA and multiplexed quantitative PCR as benchmarks to systematically evaluate the sensitivity and accuracy of various single-cell RNRNA-seq approaches. We show that single-cell RNRNA-seq can be used to perform accurate quantitative transcriptome measurement in individual cells with a relatively small number of sequencing reads and that sequencing large numbers of single cells can recapitulate bulk transcriptome complexity.

引用

页码：41 / +

页数：8

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