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 screening
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.
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 selection
Cloning, PCR Cloning
Shipping Condition: Dry Ice
|Kit, PCR Cloning|
|Cloning, PCR Cloning|
|Zero Blunt PCR cloning kits contain linearized pCR-Blunt vector, ExpressLink T4 DNA ligase , 5X ExpressLink T4 DNA ligase buffer, control template, dNTPs, sterile water, and M13 forward and reverse primers|