Amplification of 5′ end cDNA with 'new RACE'

'New RACE' (rapid amplification of cDNA ends) PCR is a method for obtaining full-length cDNA for mRNA for which only part of the sequence is known. Starting with cellular mRNA, PCR is used to amplify regions between the known parts of the sequence and nonspecific tags at the ends of the cDNA. In 'new RACE', an anchor is ligated to the 5′ end of the mRNA before reverse transcription, resulting in the selective production of full-length 5′ cDNA ends. Although 'new RACE' can also be used to amplify 3′ ends, only the protocol for obtaining 5′ ends is presented here. This protocol can be completed in 1-3 days.