INVERSE PCR AMPLIFICATION OF RARE T-CELL RECEPTOR-DELTA MESSAGES FROMMUCOSAL BIOPSY SPECIMENS

Reverse transcription (RT) PCR is an important tool in studies of the repertoire of T cell antigen receptors (TCR). In combination with inve rse PCR, it can be used to amplify TCR transcripts whose V regions are unknown. Little is currently known regarding the delta TCR repertoire in human intestine. Here, RT inverse PCR was adapted to efficiently a mplify low abundance delta TCR sequences from human colonic mucosal bi opsy specimens. In this protocol, high yields of PCR products were obt ained by optimizing the conditions at which cDNA autoligation was perf ormed.