The use of reverse transcription polymerase chain reaction to analyse large numbers of mRNA species from a single cell
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Colleges, School and Institutes
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.
|Number of pages||5|
|Journal||Journal of Immunological Methods|
|Publication status||Published - 10 May 1996|
- RNA, Messenger, Polymerase Chain Reaction, DNA, Complementary, Base Sequence, Leukocytes, Mononuclear, Humans, Molecular Sequence Data, Transcription, Genetic