TOPO Blunt-End for Sequencing

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.

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)