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 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, (C) 1995 Academic Press, Inc.