Revolutions in Rapid Amplification of cDNA Ends: New Strategies for Polymerase Chain Reaction Cloning of Full-Length cDNA Ends

Rapid amplification of cDNA ends (RACE) is a polymerase chain reaction (PCR)-based technique which was developed to facilitate the cloning of full-length cDNA 5′- and 3′-ends after a partial cDNA sequence has been obtained by other methods, While RACE can yield complete sequences of cDNA ends in onl...

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Bibliographic Details
Published in:Analytical Biochemistry 1995-05, Vol.227 (2), p.255-273
Main Author: Schaefer, B.C.
Format: Article
Language:English
Subjects:
Base Sequence
Cloning, Molecular
DNA, Complementary - genetics
Molecular Sequence Data
Polymerase Chain Reaction - methods
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Summary:Rapid amplification of cDNA ends (RACE) is a polymerase chain reaction (PCR)-based technique which was developed to facilitate the cloning of full-length cDNA 5′- and 3′-ends after a partial cDNA sequence has been obtained by other methods, While RACE can yield complete sequences of cDNA ends in only a few days, the RACE procedure frequently results in the exclusive amplification of truncated cDNA ends, undermining efforts to generate full-length clones, Many investigators have suggested modifications to the RACE protocol to improve the effectiveness of the technique, Based on first-hand experience with RACE, a critical review of numerous published variations of the hey steps in the RACE method is presented. Also included is a detailed, effective protocol based on RNA ligase-mediated RACE/reverse ligation-mediated PCR, as well as a demonstration of its utility,
ISSN:0003-2697
1096-0309
DOI:10.1006/abio.1995.1279