Invitrogen™

TA Cloning™ Kit, Dual Promoter, with pCR™II Vector and One Shot™ TOP10F' Chemically Competent E. coli

Catalog number:  K206001

Related applications:

PCR Cloning

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Description

The TA Cloning® Kit Dual Promoter with pCR™II vector provides a quick, one-step cloning strategy for directly inserting a Taq-amplified PCR product into a plasmid vector. The T7 and Sp6 promoters of the pCR™II vector allow in vitro transcription of the insert to produce sense or anti-sense products. The TA Cloning Kit Dual Promoter uses the pCR™II 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 Dual Promoter:
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™II vector provides:
• 3'-T overhangs for direct ligation of Taq-amplified PCR products
• T7 and Sp6 promoters for in vitro RNA transcription and sequencing
• 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: with One Shot® INVF' Chemically Competent E. coli (K2050-01 and K2050-40), One Shot® TOP10F' Chemically Competent E. coli (K2060-01 and K2060-40), and without competent cells (K2750-20 and K2750-40) in 20- and 40-reaction kit sizes.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Bacterial Content: Not Included
Includes: Linearized pCRII vector, ExpressLink T4 DNA ligase, 5X ExpressLink T4 DNA ligation buffer, dNTPs, 10X PCR buffer, sterile water, controls, One Shot TOP10F' chemically competent E. coli, S.O.C. medium, and a supercoiled control plasmid.
Quantity: 20 reactions
Vector: TA Cloning Vectors
Bacterial or Yeast Strain: TOP10F´
Cloning Method: TA Cloning®
Product Line: One Shot™

Contents & storage

This kit contains linearized pCR™II vector, ExpressLink™ T4 DNA ligase, 5X ExpressLink™ T4 DNA ligation buffer, dNTPs, 10X PCR buffer, sterile water, and controls. 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.