Zero Blunt™ PCR Cloning Kit provides a simple method for efficient (>80%) cloning using correction Reads blunt-ended PCR products amplified by thermostable DNA polymerase. The Zero Blunt™ PCR Cloning Kit uses the versatile pCR™-Blunt cloning vector and ExpressLink™ T4 DNA ligase to generate a ligation product in a five-minute room temperature ligation step.

Features of Zero Blunt™ Cloning™ Kit containing pCR™-Blunt vector:
•Quick and convenient—5 minutes room temperature ligation
• Efficient—the ccdB gene for positive selection produces >80% of clones with the correct insert
• Flexible—Selectable kanamycin or Zeocin™ resistance for flexible antibiotic selection

The pCR™-Blunt vector provides:
•Flanking the PCR product insertion site EcoRI site for resectable insert
• T7 promoter/primer site for in vitro RNA transcription and sequencing
• For sequencing or M13 forward and reverse primer sites for PCR screening

How Zero Blunt™ PCR Cloning Works
The Zero Blunt™ PCR Cloning Kit is designed for use in non-recombinant Clone blunt-ended PCR fragments (or any blunt-ended DNA fragment) with low background. The pCR™-Blunt vector contains the lethalE. coliccdB gene fused to the C terminus ofLacZα (Bernard et al., 1994). Ligation of blunt-ended PCR fragments disrupts expression of thelacZα-ccdB gene fusion, allowing only positive recombinants to grow upon transformation. When the transformation mixture is plated, cells containing the non-recombinant vector are killed.

Kit Configuration
Zero Blunt™ PCR Cloning Kit is available in a variety of configurations: Contains One Shot™ TOPO10 chemically competent E. coli (K2700-20 and K2700-40) and without competent cells (K2750-20 and K2750-40), available in 20 and 40 reaction kit sizes.