Invitrogen™ pENTR™/D-TOPO™ Cloning Kit, with One Shot™ Mach1™-T1^R Chemically Competent E. coli

Use highly efficient, 5-minute cloning strategy to directionally clone blunt-end PCR product into vector for entry into Gateway System or MultiSite Gateway System

Brand:  Invitrogen™ K243520

Product Code. 10246952

Includes: Ready-to-use TOPO vector, dNTPs, salt solution, sterile water, universal M13 sequencing primers, TOP10 or Mach1-T1^R Chemically E. coli, S.O.C. Medium, and a pUC19 control plasmid.

Description

• 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™
• No. of Reactions: 20 reactions
• Product Type: TOPO Cloning Kit

• 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-T1^R 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™
• Vector: pENTR

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