Reverse Transcription and PCR

Abstract

The ability to study comparatively the expression of messenger RNA (mRNA) populations from different samples has become accessible by the novel application of reverse transcription (RT), together with the polymerase chain reaction (PCR), to isolate gene sequences which can then be visualized by displaying on a sequencing gel. Differentially expressed transcripts can be analysed by Northern blotting or semiquantitative PCR to confirm differential expression. Utilization of various primer combinations facilitates the isolation of approximately 15 000 mRNA species, which represents gene expression in a mammalian cell at any one time.

Keywords: mRNA

Figure 1.

An overnight exposure of a differential display polyacrylamide gel. RT‐PCR with an oligo dT‐primer and a random primer utilizing α‐32P‐labelled dATP. The gel was run for 2 h at 1200 V; each lane represents one reaction.

Figure 2.

The protocol for differential display.

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References

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How to Cite close
Woodgate, L, and Mills, K(Mar 2003) Reverse Transcription and PCR. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0003751]