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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.

Speed with Flexibility

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.

The pCR™-Blunt Vector Features:

  • ccdB gene for positive selection
  • Choice of kanamycin or Zeocin™ resistance for flexible antibiotic selection
  • 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
 

Performance and Value

Zero Blunt® Cloning Kits are available without competent cells (K2750-20 and K2750-40) or with One Shot® TOP10 Chemically Competent E. coli (K2700-20 and K2700-40), in both 20 and 40-reaction sizes.

 

  Zero Blunt® Cloning Kits
Cloning efficiency95% (with control)
Time for ligation5 minutes
Temperature for ligationRoom Temperature
PCR Clean-up required?No
Selection AntibioticKanamycin & Zeocin™

Kit Contents

Vector
Ligation Buffer
Forward and reverse sequencing primers
dNTPs
Sterile Water
Control DNA template

Ordering Information

Catalog # Name Size List Price (USD) Qty
K275020 Zero Blunt® PCR Cloning Kit, without competent cells 20 reactions 234.00
K275040 Zero Blunt® PCR Cloning Kit, without competent cells 40 reactions 390.00
K270020 Zero Blunt® PCR Cloning Kit 20 reactions 439.00
K270040 Zero Blunt® PCR Cloning Kit 40 reactions 808.00

For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.