CloneJET PCR Cloning Kit is an advanced positive selection system for high-efficiency cloning of PCR products generated with any thermostable DNA polymerase. Any other blunt or sticky-end DNA fragment can be cloned. It is ideal for phosphorylated or non-phosphorylated DNA fragments. Ligation into the included positive selection vector takes only 5 minutes, yielding more than 99% recombinant clones. Blunt-ended PCR products generated with a proofreading enzyme are ligated directly into the cloning vector.
PCR products generated either with non-proofreading DNA polymerases or mixtures of DNA polymerases are blunted prior to ligation in 5 minutes with the thermostable DNA Blunting Enzyme provided with the kit. All common laboratory E. coli strains can be directly transformed with the ligation product.
The CloneJET PCR Cloning Kit contains a novel, ready-to-use positive selection cloning vector pJET1.2/blunt. The vector contains a lethal restriction enzyme gene that is disrupted by ligation of a DNA insert into the cloning site. As a result, only bacterial cells with recombinant plasmids are able to form colonies. Recircularized pJET1.2/blunt vector molecules lacking an insert express a lethal restriction enzyme, which kills the host E. coli cell after transformation. This positive selection drastically accelerates the process of colony screening and eliminates additional costs required for blue/white selection.
For convenience in mapping and manipulation of the insert, the pJET1.2/blunt cloning vector multiple cloning site contains two BglII recognition sequences that flank the insertion site. In addition, the vector contains a T7 promoter for in vitro and in vivo transcription as well as sequencing of the insert.
- Fast - PCR cloning in only 5 minutes
- Highest efficiency - > 99% of positive clones
- No cloning background - positive selection vector
- Versatile - ideal for blunt-end or sticky-end cloning
- Economical - no expensive blue/white screening
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In certain countries use of this product is covered by patents. Purchase of product in these countries includes non-transferable, limited license for using only this amount of product for the purchaser's own internal research. This product is licensed under one or more U.S. Patents Nos. 5,500,363 and 5,352,778 or corresponding foreign patents.
This product or the use of this product is covered by US patent application US20090042249A1 and corresponding counterparts owned by Fermentas. The purchase of this product includes a non-transferable license to use this product for the purchaser's internal research. All other commercial uses of this product, including without limitation product use for diagnostic purposes, resale of product in the original or any modified form or product use in providing commercial services require a separate license from Fermentas.
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