Use of Inverse PCR to Clone cDNA Ends

Since the first report on complementary DNA (cDNA) cloning in 1972 (1 ), the technology has been developed into a powerful and universal tool in the isolation, characterization, and analysis of both eukaryotic and prokaryotic genes. However, the conventional methods of cDNA cloning require much effort to generate a cDNA library and then screen for a large number of recombinant phages or plasmid clones. There are three major limitations in these methods. First, a substantial amount of purified mRNA (at least 1 μg) is needed as starting material to generate libraries of sufficient diversity (2 ). Second, the intrinsic difficulty of multiple sequential enzymatic reactions required for cDNA cloning often leads to low yields and/or truncated clones (3 ). Finally, screening of a library with hybridization technique is time-consuming.