| Molecular Cloning | |||||||||||||||
| CloneJET * Fast - PCR or any DNA fragments cloning in only 5 minutes | |||||||||||||||
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The CloneJET PCR Cloning Kit is an advanced positive selection system for high efficiency cloning of PCR products generated with any thermostable DNA polymerase. Additionally, any other blunt or sticky-end DNA fragment can be cloned. The kit is ideal for phosphorylated or non-phosphorylated DNA fragments. Ligation into the included positive selection vector takes only 5 minutes and yields more than 99% recombinant clones. Blunt-ended PCR products generated with a proofreading enzyme (e.g., Pfu polymerase) are ligated directly into the cloning vector. PCR products generated either with non-proofreading DNA polymerases (e.g., Taq polymerase) 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 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. Ordering information
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