5-Minute cloning and positive selection with Zero Blunt TOPO

Zero Blunt™ TOPO™ technology provides a highly efficient, 5-minute, one-step cloning strategy for the direct insertion of blunt-ended PCR products amplified with proofreading thermostable polymerases into a plasmid vector. Each kit has the Zero Blunt TOPO vector (see figure) containing the ccdB gene for positive selection, only permitting growth of plasmid vectors with recombinants so you can obtain up to 95% clones with correct insert.

The pCR Blunt II-TOPO vector is designed to clone blunt-ended PCR products generated by thermostable proofreading polymerases such as Platinum SuperFi DNA Polymerase.

Order now ›

The pCR Blunt II-TOPO vector features:

  • Contains the ccdB gene for positive selection
  • EcoRI sites flanking the PCR product insertion site for easy excision of inserts
  • Kanamycin and Zeocin™ resistance genes for your choice of selection in E. coli
  • SP6 promoter/primer site for in vitro RNA transcription and sequencing
  • M13 forward and reverse primer sites for sequencing or PCR screening

Performance and value

Each Zero Blunt TOPO for subcloning Kit contains linearized and topoisomerase I-activated pCR Blunt II-TOPOVector, Salt Solution, dNTPs, Control Template and Primers, M13 Forward and Reverse Primers, One Shot Chemically Competent or Electrocomp™ E. coli, S.O.C. Medium, and a supercoiled control plasmid. The Zero Blunt TOPO PCR Cloning Kit for Subcloning, without competent cells allows the flexibility of using your own strain for increased flexibility so competent cells and S.O.C. Medium are not provided.

In addition, a Zero Blunt TOPO Cloning Kit is available combined with a PureLink™ Quick Plasmid Miniprep Kit (50 preps) for fast plasmid purification of your TOPO-cloned inserts for downstream analysis.

Zero Blunt TOPO PCR Cloning Kits for subclobing (with pCR Blunt II TOPO vector)

Includes One Shot Competent E. coli Advantage Rxn number Cat. No.  
None
  • Flexibility to use your own competent cell strain
10 451245 Order now
TOP10 (genotype)
  • Classic TOP10 strain
10 K2800J10 Order now
None
  • Flexibility to use your own competent cell strain
25 450245 Order now
TOP10 (genotype)
  • Classic TOP10 strain
  • Most popular kit
25 K280020 Order now
MAX Efficiency DH5 α™T1 Phage Resistant (genotype)
  • Classic DH5 cloning strain
25 K282020 Order now
TOP10 Electrocomp™ (genotype)
  • Electrocompetent cells for highest transformation efficiency
25 K286020 Order now
Mach1™T1R (genotype)
  • Fast growing cells for fast miniprep
25 K283020 Order now
TOP10 (genotype)
  • Classic TOP10 strain
  • Includes PureLink™ Quick Plasmid Miniprep Kit
25 K280002 Order now
TOP10 (genotype)
  • Classic TOP10 strain
  • Most popular kit
50 K280040 Order now
MAX Efficiency DH5 α™T1 Phage Resistant (genotype)
  • Classic DH5 cloning strain
50 K282040 Order now
TOP10 Electrocomp™ (genotype)
  • Electrocompetent cells for highest transformation efficiency
50 K286040 Order now