RNA LIGASE-MEDIATED RACE: AN EFFECTIVE METHOD FOR THE CLONING OF FULL-LENGTH cDNAENDS

Rapid Amplification of cDNAEnds (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. In this chapter, the major variations of the RACE method are briefly reviewed, with particular emphasis on RNA Ligase-Mediated RACE (RLM-RACE) which offers a number of advantages over other RACE techniques. A detailed pro-tocol for RNA Ligase-Mediated RACE is presented.