[shortdesc]Topo isomerase based fast zero background cloning kit for blunt DNA fragments. Both Amp^R and Kan^R 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-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.

Fast: Instant transformation after mixing PCR fragment with vector, no incubation needeed.

CloneSmarter™ Zero Background Blunt Topoisomerase Cloning Kit Flow Chart:

[/els_overview]

[els_components]

	Kit Size
Components	C5861-25/C5862-25 (25 rxns)	C5861-50/C5862-50 (50 rxns)
pCloneEZ-Blunt-Amp/HC or pCloneEZ-Blunt-Kan/HC (20 ng/μl)	25 µl	50 µl
10X Enhancer	30 µl	60 µl
Control Insert lacZα (20ng/μl)	10 µl	10 µl
Blank Control Vector (100ng/μl)	20 µl	20 µl
M13F Sequencing Primer (10pmol)	100 µl	100 µl
M13R Sequencing Primer (10pmol)	100 µl	100 µl

[/els_components]

[els_protocol]

CloneSmarter™ Zero Background Topo Cloning Protocol:

[/els_protocol]

//here goes vector information

[additional]

Vector Sequence Files:

Snapgene File	Genbank File
pCloneEZ-Blunt-Amp-HC	pCloneEZ-Blunt-Amp-HC
pCloneEZ-Blunt-Kan-HC	pCloneEZ-Blunt-Kan-HC

Key Features:

[/additional]

//Here ends vector information

[els_citations]Click the links below to learn about recent publications citing our products from Google Scholar:

Epoch Biolabs (our old name)

Epoch Life Science (our correct name)

Epoch Life Sciences (typo by some authors)

[/els_citations]

The table header
The table body	with two columns

CloneSmarter™ Zero Background Topo Blunt DNA Cloning Kit