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 Superfi) 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.
The pCR 4 Blunt-TOPO vector

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. coliAdvantageReactionsCat. No. 
None
  • Flexibility to use your own competent cell strain
10450159Order now
TOP10 (genotype)
  • Classic TOP10 strain
  • Most popular kit
10K2875J10Order now
TOP10 (genotype)
  • Classic TOP10 strain
  • Most popular kit
25K287520Order now
TOP10 Electrocomp (genotype)
  • Electrocompetent cells for highest transformation efficiency
25K288020Order now
MAX Efficiency DH5α T1 Phage (genotype)
  • Classic DH5 cloning strain
25K289520Order now
Mach1 (genotype)
  • Fast growing cells for fast miniprep
25K283520Order now
TOP10 (genotype)
  • Classic TOP10 strain
  • Most popular kit
50K287540Order now
TOP10 Electrocomp (genotype)
  • Electrocompetent cells for highest transformation efficiency
50K288040Order now