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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
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
| Directional TOPO™ | |
| 20 reactions | |
| TOPO Cloning Kit | |
| 20 Reactions |
| 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. | |
| One Shot™ | |
| N-Terminal | |
| pENTR |
For Research Use Only. Not for use in diagnostic procedures.