title: The use of reverse transcription polymerase chain reaction to analyse large numbers of mRNA species from a single cell.
abstract: 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.
authors.affiliation: Department of Immunology and Cell Biology, Forschungsinstitut Borstel, Germany.
authors.forename: K M
authors.initials: KM
authors.lastname: Toellner
authors.forename: D
authors.initials: D
authors.lastname: Scheel-Toellner
authors.forename: U
authors.initials: U
authors.lastname: Seitzer
authors.forename: R
authors.initials: R
authors.lastname: Sprenger
authors.forename: L
authors.initials: L
authors.lastname: Trümper
authors.forename: C
authors.initials: C
authors.lastname: Schlüter
authors.forename: H D
authors.initials: HD
authors.lastname: Flad
authors.forename: J
authors.initials: J
authors.lastname: Gerdes
mesh_terms: 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
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger