Rapid cDNA Cloning by PCR Screening (RC-PCR)

Now that the draft sequence of the human genome is available (1 ,2 ), cDNA cloning based on its sequence from a library is no longer an experimental goal, but a starting point and a routine laboratory practice. Hybridization screening with either radio-labeled or nonradio-labeled probes had been commonly used for cDNA cloning before, but it is laborious and time-consuming in the post-genome era. Application of the polymerase chain reaction (PCR) is surprisingly widespread since its discovery (3 ). Here an application to cDNA library screening is reported: rapid cloning of full-length cDNAs by screening pools of cDNAs by PCR (RC-PCR) (4 ). This PCR-based cDNA screening technique is applicable to both bacteria and phage libraries.