Methods for qPCR gene expression profiling applied to 1440 lymphoblastoid single cells

被引：117

作者：

Livak, Kenneth J. ^{[1
]}

Wills, Quin F. ^{[2
]}

Tipping, Alex J. ^{[3
]}

Datta, Krishnalekha ^{[1
]}

Mittal, Rowena ^{[1
]}

Goldson, Andrew J. ^{[4
]}

Sexton, Darren W. ^{[5
]}

Holmes, Chris C. ^{[2
]}

机构：

[1] Fluidigm Corp, San Francisco, CA 94080 USA

[2] Univ Oxford, Dept Stat, Oxford OX1 3TG, England

[3] UCL, UCL Canc Inst, Stem Cell Lab, London WC1E 6BT, England

[4] Univ E Anglia, Sch Biol Sci, Biomed Res Ctr, UEA Flow Cytometry Serv, Norwich NR4 7TJ, Norfolk, England

[5] Univ E Anglia, Norwich Med Sch, Biomed Res Ctr, Norwich NR4 7TJ, Norfolk, England

来源：

METHODS | 2013年 / 59卷 / 01期

基金：

英国医学研究理事会;

关键词：

Single-cell gene expression profiling; High throughput qPCR; Real-time PCR; Microfluidic arrays; Eukaryotic transcription; Stochastic noise in gene expression; MESSENGER-RNA; TRANSCRIPTION; QUANTIFICATION;

D O I：

10.1016/j.ymeth.2012.10.004

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：71 / 79

页数：9

共 12 条

[1] Gene expression profiling in single cells from the pancreatic islets of Langerhans reveals lognormal distribution of mRNA levels [J].