THE USE OF REVERSE TRANSCRIPTION-POLYMERASE CHAIN-REACTION TO ANALYZELARGE NUMBERS OF MESSENGER-RNA SPECIES FROM A SINGLE-CELL

A PCR method is described for determining the expression of multiple h eterogeneous mRNAs from single cells. The total mRNA pool of a single selected cell is subjected to reverse transcription and subsequent tai ling with poly(dA). This cDNA is preamplified by a sequence non-specif ic PCR protocol using oligo(dT)-containing primers. The single cell cD NA library obtained permits the analysis of virtually unlimited number s of mRNA species per cell using sequence-specific PCR. This procedure of multiple mRNA analysis enables phenotyping of any cell for its mRN A composition and could be used to study the cytokine mRNA expression of individual human T cells ex vivo. The method should greatly facilit ate the analysis of combinatorial expression of known genes in any cel l.