title: Construction of a High Efficiency PCR Products Cloning T Vector Using pGEM-5zf (+).
abstract: A highly efficient cloning vector was constructed for cloning PCR products by inserting an 80 bp DNA fragment into pGEM-5zf (+) vector. The Xcm I digestion of this vector gave rise to a 3' overhanging deoxythymidine offering the possibility of cloning PCR products with 3' adenosine overhang created by Taq DNA polymerase. Furthermore, two EcoR I sites were added to the construct for identification of recombinant plasmids using a single restriction enzyme. Taken together, the more efficient cloning performance and the lower cost of this vector as compared to the commercial T vector, suggests that it may be one of the best T vectors for cloning of PCR products.
authors: Division of Clinical Immunology, Department of Laboratory Medicine, Karolinska University Hospital Huddinge, SE-141 86, Stockholm, Sweden.
authors: Yaofeng
authors: Y
authors: Zhao
authors: Zhancai
authors: Z
authors: Liu
authors: Shuyang
authors: S
authors: Yu
authors: Sicheng
authors: S
authors: Wen
authors: Lennart
authors: L
authors: Hammarstrom
authors: Hodjattallah
authors: H
authors: Rabbani
keywords: Cloning; PCR products; T vector; pGEM-5zf(+)