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 (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.

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 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 and blue⁄white screening.
  • 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.
 

pCR®-4 Blunt TOPO® vector.

The pCR™4 Blunt-TOPO® vector

Performance and value

Each Zero Blunt® TOPO® for subcloning® Kit 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.

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克隆的 PCR 产物的应用 产品 Includes One Shot® Competent E. coli 优点 Rxn number 目录号  
测序 Zero Blunt® TOPO® PCR Cloning Kit (with pCR®4 Blunt TOPO) TOP10 (genotype)
  • Classic TOP10 strain
  • Most popular kit
10 K2875J10 马上订购
TOP10 (genotype)
  • Classic TOP10 strain
  • Most popular kit
20 K287520 立即订购
TOP10 Electrocomp™ (genotype)
  • Electrocompetent cells for highest transformation efficiency
20 K288020 立即订购
MAX Efficiency® DH5 α ™T1 Phage (genotype)
  • Classic DH5 cloning strain
20 K289520 马上订购
Mach I™ (genotype)
  • 为生长快速的细胞,利于进行快速地质粒的小量提取
20 K283520 马上订购
TOP10 (genotype)
  • Classic TOP10 strain
  • Most popular kit
40 K287540 马上订购
TOP10 Electrocomp™ (genotype)
  • Electrocompetent cells for highest transformation efficiency
40 K288040 马上订购