The use of reverse transcription polymerase chain reaction to analyse large numbers of mRNA species from a single cell

Research output: Contribution to journalArticle

Authors

  • U Seitzer
  • R Sprenger
  • L Trümper
  • C Schlüter
  • H D Flad
  • J Gerdes

Colleges, School and Institutes

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.

Details

Original languageEnglish
Pages (from-to)71-5
Number of pages5
JournalJournal of Immunological Methods
Volume191
Issue number1
Publication statusPublished - 10 May 1996

Keywords

  • RNA, Messenger, Polymerase Chain Reaction, DNA, Complementary, Base Sequence, Leukocytes, Mononuclear, Humans, Molecular Sequence Data, Transcription, Genetic