Comprehensive qPCR profiling of gene expression in single neuronal cells

被引：120

作者：

Citri, Ami ^{[1
]}

Pang, Zhiping P. ^{[2
,3
]}

Suedhof, Thomas C. ^{[2
,4
]}

Wernig, Marius ^{[5
]}

Malenka, Robert C. ^{[1
]}

机构：

[1] Stanford Univ, Dept Psychiat & Behav Sci, Sch Med, Stanford, CA 94305 USA

[2] Stanford Univ, Dept Mol & Cellular Physiol, Sch Med, Stanford, CA 94305 USA

[3] Univ Med & Dent New Jersey, Robert Wood Johnson Med Sch, Child Hlth Inst New Jersey, Dept Neurosci & Cell Biol, New Brunswick, NJ USA

[4] Howard Hughes Med Inst, Chevy Chase, MD USA

[5] Stanford Univ, Dept Pathol, Inst Stem Cell Biol & Regenerat Med, Sch Med, Stanford, CA 94305 USA

来源：

NATURE PROTOCOLS | 2012年 / 7卷 / 01期

关键词：

REAL-TIME PCR; DIVERSITY;

D O I：

10.1038/nprot.2011.430

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A major challenge in neuronal stem cell biology lies in characterization of lineage-specific reprogrammed human neuronal cells, a process that necessitates the use of an assay sensitive to the single-cell level. Single-cell gene profiling can provide definitive evidence regarding the conversion of one cell type into another at a high level of resolution. The protocol we describe uses Fluidigm Biomark dynamic arrays for high-throughput expression profiling from single neuronal cells, assaying up to 96 independent samples with up to 96 quantitative PCR (qPCR) probes (equivalent to 9,216 reactions) in a single experiment, which can be completed within 2-3 d. The protocol enables simple and cost-effective profiling of several hundred transcripts from a single cell, and it could have numerous utilities.

引用

页码：118 / 127

页数：10

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