Thermo Scientific™ CloneJET™ PCR Cloning Kit

Efficiently clone PCR products generated with any thermostable DNA polymerase using Thermo Scientificâ„¢ CloneJETâ„¢ PCR Cloning Kit, a positive selection system.

Overview
Brand: Thermo Scientific™

Manufacturer Part Number: K1232A

1 ML GENEBLAZER(R) D2 GQO5 NFAT BLA CHO K1 CELLline 1mL

UNSPSC: 12352200

Code: 50

Additional Details:
Additional Details: Weight: 0.50000kg



Disclaimers: 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.

Product Code. 10507533

Quantity Price
1 £ 7950.0 / 40 reactions
EU Stock 65
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For Research Use Only. All usage must comply with product instructions.
 

Description and Specification

Specification

For Use With (Application) 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
Product Type CloneJETâ„¢ PCR Cloning Kit
Quantity 40 reactions

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.

  • 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

Recommended for:

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

Note:

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.