CHANGYUZeroBackgroundpTOPO-TACloningKit

CHANGYU pTOPO-Blunt Kit

Product Manual 

 

Cat# Size Price ($) Bulk order
CYCV1601 20 apps 89 Contact
CYCV1602 80 apps 309 Contact

 

Storage

-20°C, stable for 12 months.

 

Description

The 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. 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 pTOPO-Blunt included in this kit are high copy number plasmids engineered to tolerate mild toxic genes. Regions flanking the cloning site of pTOPO-Blunt vectors have multiple common restriction sites for release of the cloned fragment by single or double restriction digestion.

 

Features

1. Whether blunt end or A-end PCR product can complete the connection instantly; 

2. Without ice bath and heat shock, complete transformation within 5 minutes at RT; 

3. From the connection to the entire coating plates only 20 minutes;

4. No self-connection, zero background, no blue white screening, nearly 100% positive;

5. Can connect up to 10kb fragments, is the easiest, fastest TOPO vector;

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