The use of reverse transcription polymerase chain reaction to analyse large numbers of mRNA species from a single cell.
A PCR method is described for determining the expression of multiple heterogeneous mRNAs from single cells. The total mRNA pool of a single selected cell is subjected to reverse transcription and subsequent tailing with poly(dA). This cDNA is preamplified by a sequence non-specific PCR protocol using oligo(dT)-containing primers. The single cell cDNA library obtained permits the analysis of virtually unlimited numbers of mRNA species per cell using sequence-specific PCR. This procedure of multiple mRNA analysis enables phenotyping of any cell for its mRNA composition and could be used to study the cytokine mRNA expression of individual human T cells ex vivo. The method should greatly facilitate the analysis of combinatorial expression of known genes in any cell.
Toellner
K M
KM
Department of Immunology and Cell Biology, Forschungsinstitut Borstel, Germany.
Scheel-Toellner
D
D
Seitzer
U
U
Sprenger
R
R
Trümper
L
L
Schlüter
C
C
Flad
H D
HD
Gerdes
J
J
D001483:Base Sequence; D018076:DNA, Complementary / Q000032:analysis; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000737:chemistry; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis*; D014158:Transcription, Genetic / Q000276:immunology
D018076:DNA, Complementary; D012333:RNA, Messenger