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Updated: May 8, 2026

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning
Published on: February 5, 2021
An efficient method to prepare PCR cloning vectors
Soon Gyu Hong1, Ji Young Choi, Barry M Pryor
1Polar BioCenter, Korea Polar Research Institute, KORDI, Songdo Techno Park, Songdo-dong 7-50, Yeonsu-gu, Incheon 406-840, Korea. ; Division of Plant Pathology and Microbiology, Department of Plant Sciences, College of Agriculture and Life Sciences, University of Arizona, Tucson, Arizona 85721, USA.
Abstract:
An improved procedure for preparing PCR cloning vectors was developed. This procedure includes the incorporation of adapters to create XcmI restriction enzyme sites in pBluescript II SK(+) vectors, digestion with XcmI followed by further digestion of the small fragment produced by XcmI digestion with additional enzymes, and purification with PCR purification kits. Using this procedure, PCR cloning vectors with high ligation efficiencies and low blue or false-positive colonies were obtained.

