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

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

Supplier:  Invitrogen™ K240020

Catalog No. K240020

Includes: pENTR/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 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
• Directional cloning vector for entry to Gateway System
• 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 the choice of Gateway Destination vectors
• The pENTR/D-TOPO vector includes 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 the choice of attR containing Gateway destination vectors
• Kanamycin resistance gene and a 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
• Circumvents the limitations of restriction mediated cloning, enabling to access virtually any expression system in a simple one hour, 99%-efficient and reversible, Gateway recombination reaction
• Move same sequence of DNA between different vectors without using restriction enzymes, ligase, subcloning steps, screening of countless colonies, or resequencing to save time, money, and effort

Cloning, Gateway Cloning

Specifications

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

• Content And Storage: pENTR™⁄D-TOPO™ Cloning Kit contains pENTR⁄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™
• Vector: pENTR

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