The TA Cloning® Kit with pCR™2.1 vector provides a quick, one-step cloning strategy for directly inserting a Taq-amplified PCR product into a plasmid vector. The TA Cloning® Kit uses the pCR™2.1 cloning vector and ExpressLink™ T4 DNA Ligase to generate a ligation product in a fifteen-minute, room-temperature ligation step. Reactions typically yield >80% recombinants containing inserts.
Features of the TA Cloning® Kit with pCR™2.1 vector: • Fast & convenient—15-minute, room-temperature ligation • Efficient—blue/white screening and >80% clones with correct insert • Flexible—choice of kanamycin or ampicillin resistance for flexible antibiotic selection • Hassle-free—eliminates any enzymatic modifications of the PCR product • Streamlined—does not require the use of PCR primers that contain restriction sites
The pCR™2.1 vector provides: • 3'-T overhangs for direct ligation of Taq-amplified PCR products • T7 promoter for in vitro RNA transcription and sequencing • A versatile polylinker with flanking EcoR I sites for easy excision of inserts • M13 forward and reverse primer sites for sequencing
How TA Cloning® Works Taq polymerase has a non-template-dependent activity that adds a single deoxyadenosine (A) to the 3' ends of PCR products. The linearized vector supplied in this kit has single 3' deoxythymidine (T) residues. This allows PCR inserts to ligate efficiently with the vector.
Kit Configurations The TA Cloning® Kit is offered in a variety of configurations: without competent cells (K2020-20 and K2020-40), with One Shot® INVF' Chemically Competent E. coli (K2000-01 and K2000-40), with One Shot® TOP10F' Chemically Competent E. coli (K2030-01 and K2030-40), and with One Shot® TOP10 Chemically Competent E. coli (K2040-01 and K2040-40) in 20- and 40-reaction kit sizes.
For Research Use Only. Not for use in diagnostic procedures.
TA Cloning® kits contain linearized pCR™2.1 vector, ExpressLink™ T4 DNA ligase, 5X ExpressLink™ T4 DNA ligation buffer, dNTPs, 10X PCR buffer, sterile water, and controls. The competent cell kits contain One Shot® chemically competent E. coli, S.O.C. medium, and a supercoiled control plasmid.
Store One Shot® E. coli at -80°C. Store all other components at -20°C. All reagents are guaranteed stable for 6 months when properly stored.