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® Pfx DNA Polymerase.

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 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

pCR®Blunt II-TOPO® vector diagram

The pCR®Blunt II-TOPO® vector

Performance and value

Each Zero Blunt® TOPO® for subcloning® Kit contains linearized and topoisomerase I-activated pCR®Blunt II-TOPO® Vector, 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.

Application of cloned PCR product Product Includes One Shot® Competent E. coli Advantage Rxn number Cat. No.  
General subcloning Zero Blunt® TOPO® PCR Cloning Kit (with pCR® Blunt II TOPO® vector) 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