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Invitrogen™ pENTR™/D-TOPO™ Cloning Kit, with One Shot™ Mach1™-T1R Chemically Competent E. coli
Includes
Ready-to-use TOPO vector, dNTPs, salt solution, sterile water, universal M13 sequencing primers, TOP10 or Mach1-T1R Chemically E. coli, S.O.C. Medium, and a pUC19 control plasmid.
- Blunt-end PCR products clone directionally at greater than 90% efficiency, with no ligase, post-PCR procedures, or restriction enzymes required
- 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/D-TOPO vector, incubate 5 minutes, and transform provided competent E. coli cells
- Resulting attL containing Gateway Entry clones are ready for efficient recombination with choice of Gateway Destination vectors
- pENTR/D-TOPO vector includes M13 and T7 primer sequencing sites and attL recombination sites flanking PCR product insertion site., allowing clones to be easily sequence-verified and recombined into choice of attR containing Gateway destination 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 the topoisomerase ligation reaction
- Move same sequence of DNA between different vectors without using restriction enzymes, ligase, subcloning steps, screening of countless colonies, or resequencing
Cloning/Gateway Cloning
Specifications
Cloning Method
Directional TOPO™
Content And Storage
Each Directional TOPO™ pENTR Cloning Kit contains the appropriate, ready-to-use TOPO™ vector, dNTPs, salt solution, sterile water, universal M13 sequencing primers, TOP10 or Mach1-T1R 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.
No. of Reactions
20 reactions
Product Line
One Shot™
Product Type
TOPO Cloning Kit
Vector
pENTR
For Research Use Only. Not for use in diagnostic procedures.
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