Thermo Fisher, K263520, pENTR™/SD/D-TOPO™ Cloning Kit, with One Shot™ Mach1™-T1^R Chemically Competent <i>E. coli</i>

Quantity: 20 Reactions
Unit Size: Each
• Blunt-end PCR products clone directionally into pENTR™/SD/D-TOPO™ entry vector at greater than 90% efficiency, and include upstream Shine-Dalgarno sequence for expression in prokaryotes
• 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 SD/SD D-TOPO vector, incubate 5 minutes and transform provided competentE. colicells
• Resulting attL containing clones are ready for efficient recombination with your choice of vectors
• pENTR™ /SD SD/SD D-TOPO™ vector includes T7 gene 10 translational enhancer and ribosome binding site (RBS) for optimal expression of native protein after recombination with prokaryotic vectors
• Also use vector for expression in other hosts systems by recombination with different destination vectors
• 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 attR containing destination vectors
• A Kanamycin resistance gene and a pUC origin are used for selection and high-copy propagation inE. coli
• No PCR clean up, vector preparation, or other time-intensive DNA manipulation steps
• Four base overhang on vector pairs with four base sequence designed into forward primer used inthe PCR reaction to provide directionality to topoisomerase ligation reactionCloning, Gateway Cloning