Description
Quantity: 10 Reactions
Unit Size: Each
• Each kit has the Zero Blunt™ TOPO™ vector containing the ccdB gene for positive selection, only permitting growth of plasmid vectors with recombinants
• Available with a variety of competent cells, or without competent cells
• Fast and easy—go from PCR to clones in just 3 steps, including a 5-minute cloning reaction
• Efficient—obtain up to 95% clones with correct insert
• Proven—enabling reliable performance for over a decade with over 20,000 citations
• Simple —no ligase, post-PCR procedures, or PCR primers containing specific sequences required
• Accurate—for use with high fidelity DNA polymerases, no need for A-overhang on PCR product
• The pCR™Blunt II-TOPO™ vector is designed for cloning of blunt-ended PCR products generated by thermostable proofreading polymerases such as Platinum™ Pfx DNA Polymerase.
• Allows for direct selection of recombinants via disruption of the lethalE. coligene ccdB. The vector contains the ccdB gene fused to the C-terminus of the LacZα fragment. Ligation of a blunt-end PCR product disrupts expression of the LacZα-ccdB gene fusion, permitting growth of only positive recombinants upon transformation. Cells that contain non-recombinant vector are killed upon plating. Therefore, blue/white screening is not required.
• Using TOPO™ cloning technology, there is no need for PCR primers containing specific sequences, post-PCR procedures, vector preparation, or other time-intensive DNA manipulation steps. Just add your PCR reaction straight to the provided topoisomerase-charged vector, incubate 5 minutes, and transformE. colicompetent cells.
• Zero Blunt™ TOPO™ PCR Cloning kits for direct insertion of blunt-ended PCR products into a plasmid vector can be purchased with a variety of competent cells that deliver different advantages
