pGEM® T Vector Systems, Promega
Supplier: Promega Corporation
The pGEM-T Vector Systems are convenient systems for cloning PCR products.
- Convenience for Cloning PCR Products
- Insert excision with a BstZI single digest
- Ligation can be completed in 1 hour at room temperature
- Available with or without competent cells
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.
The pGEM-T Vector System II contains JM109 Competent Cells in addition to all of the pGEM-T Vector System I components.
This single T overhangs at the insertion site to 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 to the 3´ ends of amplified fragments. The multiple cloning site is flanked by recognition sites for the restriction enzyme BstZI, which allows the release of the insert by a single-enzyme digestion. Alternatively, a double digestion may be used to release the insert from the vector.
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