An Efficient Method to Prepare PCR Cloning Vectors

Abstract

An improved procedure for preparing PCR cloning vectors was developed. This procedure includes the incorporation ofadapters to create XcmI restriction enzyme sites in pBluescript II SK(+) vectors, digestion with XcmI followed by furtherdigestion of the small fragment produced by XcmI digestion with additional enzymes, and purification with PCR purificationkits. Using this procedure, PCR cloning vectors with high ligation efficiencies and low blue or false-positive colonies wereobtained.