Invitrogen™ pENTR™/TEV/D-TOPO™ Cloning Kit, with One Shot™ TOP10 Chemically Competent E. coli

Use highly efficient, 5-minute cloning strategy to directionally clone blunt-end PCR product into vector

Supplier:  Invitrogen™ K252520

Catalog No. K252520

Includes: pENTR/TEV/D-TOPO vector, dNTPs, salt solution, sterile water, universal M13 sequencing primers, OneShot TOP10 Chemically E. coli, S.O.C. Medium, and a pUC19 control plasmid.

Description

• Blunt-end PCR products clone directionally into pENTR/TEV/D-TOPO entry vector at greater than 90% efficiency, and include TEV protease-dependent cleavage site for removal of N-terminal tags from expressed proteins
• Rapidly shuttle cloned genes between multiple vector systems
• Reliable performance
• For access to Gateway System, PCR amplify gene of interest and add product straight to provided topoisomerase charged pENTR/SD/D-TOPO vector, incubate 5 minutes, and transform provided competent E. coli cells
• Resulting attL containing Gateway Entry clones are ready for efficient recombination
• Optimized pENTR/TEV/D-TOPO Vector includes Tobacco Etch Virus (TEV) recognition site for protease-dependent cleavage of N-terminal tag from recombinant protein
• Vector has M13 and T7 primer sequencing sites and attL recombination sites flanking the PCR product insertion site, allowing clones to be easily sequence-verified and recombined into choice of vectors
• Kanamycin resistance gene and pUC origin are used for selection and high-copy propagation in E. coli
• Four base overhang on vector pairs with four base sequence designed into forward primer used in PCR reaction to provide directionality to topoisomerase ligation reaction

Cloning, Gateway Cloning

Specifications

• Cloning Method: Directional TOPO™
• No. of Reactions: 20 reactions
• Product Type: TOPO Cloning Kit
• Vector: pENTR

• Content And Storage: pENTR™⁄TEV⁄D-TOPO™ Cloning Kit contains pENTR⁄TEV⁄D-TOPO™ vector, dNTPs, salt solution, sterile water, universal M13 sequencing primers, OneShot™ TOP10 Chemically E. coli, S.O.C. Medium, and a pUC19 control plasmid. Store Competent E. coli at -80°C. Store all other components at -20°C.
• Product Line: One Shot™
• Protein Tag: N-Terminal

For Research Use Only. Not for use in diagnostic procedures.