pGEM-T Vector System II
The pGEM-T Vector Systems are convenient systems to clone PCR products. The pGEM-T Vector is prepared by cutting the pGEM-5Zf(+) Vector with EcoRV and adding a 3' terminal thymidine to both ends. These single 3'-T overhangs at the insertion site greatly improve the efficiency of ligation of a PCR product into the plasmid by preventing recircularization of the vector and providing a compatible overhang for ligation of PCR products generated by thermostable polymerases that add a single deoxyadenosine, in a template-independent fashion, to the 3'-ends of amplified fragments. The multiple cloning site is flanked by recognition sites for the restriction enzyme BstZI, allowing release of the insert by a single-enzyme digestion. Alternatively, a double digestion may be used to release the insert from the vector. The pGEM-T Vector System II contains JM109 Competent Cells in addition to all of the pGEM-T Vector System I components.