Reverse Transcription and PCR

L Woodgate, University of Cardiff, Cardiff, UK
K Mills, University of Cardiff, Cardiff, UK

Published online: March 2003

DOI: 10.1038/npg.els.0003751

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 -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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Related Sub-Topics:

Replication and Recombination | Sample Preparation in Structural Biology | Techniques and Tools in Molecular Biology | Enzymes: Structure and Action Mechanism | Nucleic Acids: Structure, Function, Metabolism
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Article Title: Reverse Transcription and PCR
 
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]