Thermo Scientific™ CloneJET™ PCR Cloning Kit with DH10B Competent Cells, 40 Reactions

Thermo Scientific™ CloneJET™ PCR Cloning Kit with DH10B Competent Cells, 40 Reactions

The CloneJET PCR Cloning Kit contains the 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. Re-circularized 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.

Features

• Fast—PCR cloning in only five 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

Contents

• 20 x DH10B Competent Cells for Subcloning (100 µL)
• 2 x pUC19 DNA (20 µL) (100 pg/µL)
• 2 x S.O.C. Medium (5 mL)
• pJET1.2/blunt Cloning Vector (46 µL) (50 ng/µL)
• 2X Reaction Buffer (600 µL)
• T4 DNA Ligase (24 µL) (5 U/µL)
• DNA Blunting Enzyme (46 µL)
• DNA Blunting Enzyme (46 µL)
• pJET1.2 Forward Sequencing Primer, 10 µM aqueous solution (50 µL)
• pJET1.2 Reverse Sequencing Primer, 10 µM aqueous solution (50 µL)
• Control PCR Product, 976 bp with 3’-dA overhangs (8 µL) (24 ng/µL)
• Water, nuclease-free, (1.25 mL)