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. coli ccdB gene fused to the C-terminus of LacZα.
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.
With the addition of the ExpressLink T4 DNA Ligase into Zero Blunt Cloning Kits, ligation can now be performed at room temperature in only 5 minutes with reactions typically yielding >80% recombinants containing inserts.
|Zero Blunt Cloning Kits|
|Cloning efficiency||95% (with control)|
|Time for ligation||5 minutes|
|Temperature for ligation||Room Temperature|
|PCR Clean-up required?||No|
|Selection Antibiotic||Kanamycin & Zeocin|
|Forward and reverse sequencing primers|
|Control DNA template|
For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.