CloneSmarter™ Zero Background Topo Blunt DNA Cloning Kit

CloneSmarter™ Zero Background Blunt Topoisomerase Cloning Kit is designed for fast cloning of blunt ended DNA fragments up to 10 kb generated by high fidelity DNA polymerase such as KOD, Pfu and Phusion ext. DNA fragments obtained by restriction digestion or mechanical shearing can also be cloned after end polishing to become blunt ended. It utilizes DNA strand transfer activity of Viccinia virus topoisomerase I. Vaccinia virus DNA topoisomerase I forms a 3’-phosphoryl intermediate with the plasmid vector containing cleavage recognition motif of 5’ CCCTT↓. Covalently bound topoisomerase I then transfer the incised vector DNA strand to the DNA fragment to be cloned with free 5’-OH terminuses. This transferring reaction is rapid and reproducible. The cloning vector pCloneEZ-Blunt-Amp/HC and pCloneEZ-Blunt-Kan/HC included in this kit are high copy number plasmid engineered to tolerate mild toxic genes. For fast cloning of extremely toxic genes please chose our advanced CloneRanger Topoisomerase Kit.

Does not include competent cell.
Zero Background: >90% of randomly picked clones contain the right insert.

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