5-minute cloning and positive selection with Zero Blunt TOPO vector

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 (such as Platinum Pfx DNA Polymerase) into a plasmid vector.

Each kit uses the pCR 4 Blunt-TOPO vector with specially designed sequencing primer sites that return more insert sequence and less vector sequence from each reaction. These kits include everything necessary to clone and select recombinant vectors containing your PCR fragment of choice.

Select Zero Blunt TOPO PCR Cloning Kits ›

The pCR 4 Blunt-TOPO vector features:

  • Contains the ccdB gene for positive selection
  • Short distance between sequencing primer sites and the insert site to as little as 33 bp. This means sequencing reactions give less vector sequence and more insert sequence.
  • 4 common sequencing primers: M13 forward, M13 reverse, T7, and T3
  • Both ampicillin and kanamycin resistance markers and a LacZα-ccdB gene fusion for positive selection.
  • The vector’s minimized multiple cloning site still includes flanking EcoRI sites for simplified excision of cloned PCR products and a unique Sse8387I site for generation of nested deletions prior to sequencing.
  • T7 and T3 promoters are also present for in vitro transcription.

Performance and value

Each Zero Blunt TOPO PCR Cloning Kit for Sequencing contains linearized and topoisomerase I-activated pCR 4 Blunt-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.

Zero Blunt TOPO PCR Cloning Kits for sequencing (with pCR 4-Blunt TOPO vector)

Includes One Shot competent E. coli Advantage Reactions Cat. No.  
None
  • Flexbility to use your own competent cell strain
10 450159 Order now
TOP10 (genotype)
  • Classic TOP10 strain
  • Most popular kit
10 K2875J10 Order now
TOP10 (genotype)
  • Classic TOP10 strain
  • Most popular kit
25 K287520 Order now
TOP10 Electrocomp (genotype)
  • Electrocompetent cells for highest transformation efficiency
25 K288020 Order now
MAX Efficiency® DH5α™ T1 Phage (genotype)
  • Classic DH5 cloning strain
25 K289520 Order now
Mach1™ (genotype)
  • Fast growing cells for fast miniprep
25 K283520 Order now
TOP10 (genotype)
  • Classic TOP10 strain
  • Most popular kit
50 K287540 Order now
TOP10 Electrocomp (genotype)
  • Electrocompetent cells for highest transformation efficiency
50 K288040 Order now