Thermo Fisher, K275020, Zero Blunt™ PCR Cloning Kit, without competent cells, 20 Reactions

Quantity: 20 Reactions
Unit Size: Each
The Zero Blunt™ PCR Cloning Kit uses the multipurpose cloning vector pCR™-Blunt and ExpressLink™ T4 DNA Ligase to generate a ligation product in a five-minute, room-temperature ligation step.The pCR-Blunt vector provides:
• EcoR I sites flanking the PCR product insertion site for excision of inserts
• T7 promoter/primer site for in vitro RNA transcription and sequencing
• M13 forward and reverse primer sites for sequencing or PCR screeningHow Zero Blunt PCR Cloning Works The Zero Blunt PCR Cloning Kit is designed to clone blunt PCR fragments (or any blunt DNA fragment) with a low background of non-recombinants. The pCR-Blunt vector contains the lethal E. coliccdB gene fused to the C-terminus of LacZα (Bernard et al., 1994). Ligation of a blunt PCR fragment disrupts expression of the lacZα-ccdB gene fusion permitting growth of only positive recombinants upon transformation. Cells that contain non-recombinant vector are killed when the transformation mixture is plated.Kit Configurations The Zero Blunt PCR Cloning Kit is offered in a variety of configurations: with One Shot™ TOPO10 Chemically Competent E. coli (K2700-20 and K2700-40) and without competent cells (K2750-20 and K2750-40) in 20- and 40- reaction kit sizes.
• Fast and convenient: 5-minute, room-temperature ligation
• Efficient:ccdB gene for positive selection results in >80% clones with correct insert
• Flexible: choice of kanamycin or Zeocin™ resistance for flexible antibiotic selectionCloning, PCR Cloning