Journal
METHODS
Volume 59, Issue 1, Pages 71-79Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2012.10.004
Keywords
Single-cell gene expression profiling; High throughput qPCR; Real-time PCR; Microfluidic arrays; Eukaryotic transcription; Stochastic noise in gene expression
Funding
- Medical Research Council of the United Kingdom
- Leukaemia Lymphoma Research
- EuroSyStem
- Medical Research Council [MC_UP_A390_1107] Funding Source: researchfish
- MRC [MC_UP_A390_1107] Funding Source: UKRI
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The stochastic nature of generating eukaryotic transcripts challenges conventional methods for obtaining and analyzing single-cell gene expression data. In order to address the inherent noise, detailed methods are described on how to collect data on multiple genes in a large number of single cells using microfluidic arrays. As part of a study exploring the effect of genotype on Wnt pathway activation, data were collected for 96 qPCR assays on 1440 lymphoblastoid cells. The description of methods includes preliminary data processing steps. The methods used in the collection and analysis of single-cell qPCR data are contrasted with those used in conventional qPCR. (C) 2012 Elsevier Inc. All rights reserved.
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