ABSTRACT

During the construction of a library of complementary DNA (see Profile 15), the 5-P end of the sequence (the beginning of the cDNA) is sometimes absent because the reverse transcription (which begins the synthesis from the 3-OH end) may not have been able to synthesize the cDNA completely (see Profile 27). Moreover, when cDNA is obtained by hexanucleotide random priming, one of the ends of the mRNA (5'-P or 3-OH) is absent. Thus, the information present at the ends of the molecule is lost. One technique based on the use of PCR (see Profile 24) has been implemented in order to quickly complete the ends of the mRNA. This is the RACE technique or rapid amplification of cDNA ends.