Expression profiling of single cells using 3 prime end amplification (TPEA) PCR.
The ability to relate the physiological status of individual cells to the complement of genes they express is limited by current methodological approaches for performing these analyses. We report here the development of a robust and reproducible method for amplifying 3' sequences of mRNA derived from single cells and demonstrate that the amplified cDNA, derived from individual human lymphoblastoma cells, can be used for the expression profiling of up to 40 different genes per cell. In addition, we show that 3 prime end amplification (TPEA) PCR can be used to enable the detection of both high and low abundance mRNA species in samples harvested from live neurons in rat brain slices. This procedure will facilitate the study of complex tissue function at the cellular level.
Dixon
A K
AK
The Sanger Centre, The Wellcome Genome Campus, Hinxton, Cambridge CB10 1SA, UK,Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QJ, UK.
Richardson
P J
PJ
Lee
K
K
Carter
N P
NP
Freeman
T C
TC
D000818:Animals; D002468:Cell Physiological Phenomena; D018076:DNA, Complementary / Q000032:analysis* / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics*; D051381:Rats
D018076:DNA, Complementary; D012333:RNA, Messenger