CloneSmarter™ Zero Background Topo Blunt DNA Cloning Kit

[shortdesc]Topo isomerase based fast zero background cloning kit for blunt DNA fragments. Both AmpR and KanR available for choose. Without competent cell[/shortdesc]   [els_overview]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 (refer to the illustration below). This transferring reaction is rapid and reproducible. The cloning vector pCloneEZ-B