Thermo Fisher, K270020, Zero Blunt™ PCR Cloning Kit

20 Reactions The Zero Blunt™ PCR Cloning Kit offers an easy method for high-efficiency (>80%) cloning of blunt-end PCR products amplified with proof-reading, thermostable DNA polymerases. 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. Features of the Zero Blunt™ Cloning™ Kit with pCR™-Blunt vector:
•Fast & 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 selectionThe pCR™-Blunt vector provides:
•EcoR I sites flanking the PCR product insertion site for excision of inserts
• T7 promoter/primer site forin vitroRNA transcription and sequencing
• M13 forward and reverse primer sites for sequencing or PCR screeningHow Zero Blunt™ PCR Cloning WorksThe 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 lethalE. coliccdB gene fused to the C-terminus ofLacZα (Bernard et al. , 1994). Ligation of a blunt PCR fragment disrupts expression of thelacZα-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 ConfigurationsThe Zero Blunt™ PCR Cloning Kit is offered in a variety of configurations: with One Shot™ TOPO10 Chemically CompetentE. coli(K2700-20 and K2700-40) and without competent cells (K2750-20 and K2750-40) in 20- and 40- reaction kit sizes. Bacterial or Yeast Strain: TOP10
Cloning Method: Blunt PCR
Product Line: Zero Blunt™
Product Type: PCR Cloning Kit
Promoter: T7
Quantity: 20 Reactions
Vector: Blunt DNA Cloning Vectors
Format: Kit
Unit Size: Each