Methods for qPCR gene expression profiling applied to 1440 lymphoblastoid single cells.
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.
Fluidigm Corporation, 7000 Shoreline Court, Suite 100, South San Francisco, CA 94080, USA. ken.livak@fluidigm.com
Kenneth J
KJ
Livak
Quin F
QF
Wills
Alex J
AJ
Tipping
Krishnalekha
K
Datta
Rowena
R
Mittal
Andrew J
AJ
Goldson
Darren W
DW
Sexton
Chris C
CC
Holmes
D002460:Cell Line; D003627:Data Interpretation, Statistical; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D057230:Limit of Detection; D054503:Lymphoid Progenitor Cells / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D059010:Single-Cell Analysis; D060449:Wnt Signaling Pathway