Thermo Scientific™ CloneJET™ PCR Cloning Kit
Efficiently clone PCR products generated with any thermostable DNA polymerase using Thermo Scientific™ CloneJET™ PCR Cloning Kit, a positive selection system.
Brand: Thermo Scientific™ K1232
Code : NEW
Additional Details : Weight : 2.17720kg
DescriptionAdditionally, 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.
- 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
Cloning of blunt-end or 3'-dA tailed PCR products up to 10kb; Cloning of DNA fragments generated by restriction enzymes; Sequencing of cloned DNA; In vitro and in vivo transcription of cloned inserts from the T7 promoter
Prior to electroporation always column-purify the ligation mixture using, e.g., GeneJET PCR Purification Kit, †K0701, or chloroform extract kit. Electroporation is inhibited by the presence of proteins and salts in the mixture.
|Cloning of blunt-end or 3'-dA tailed PCR products up to 10kb; Cloning of DNA fragments generated by restriction enzymes; Sequencing of cloned DNA; In vitro and in vivo transcription of cloned inserts from the T7 promoter|
|CloneJET™ PCR Cloning Kit|