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Nat Protoc. 2011 Dec 22;7(1):118-27. doi: 10.1038/nprot.2011.430.

Comprehensive qPCR profiling of gene expression in single neuronal cells.

Author information

1
Department of Psychiatry and Behavioral Sciences, Stanford University School of Medicine, California, USA. citri@stanford.edu

Abstract

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.

PMID:
22193304
PMCID:
PMC4377330
DOI:
10.1038/nprot.2011.430
[Indexed for MEDLINE]
Free PMC Article

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