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 DMA fragment into pCEM-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 DMA 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