利用TOPO快速高效地完成PCR克隆

TOPO TA subcloning kits utlize our popular pCR2.1-TOPO TA vector for the direct ligation of Taq-amplified PCR products in a 5-minute, benchtop reaction yielding up to 95% recombinants.

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 The pCR2.1-TOPO TA vector features:

  • T7 promoter and M13 forward and reverse primer sites for in vitro RNA transcription and sequencing
  • EcoR I sites flanking the PCR product insertion site for easy excision of inserts
  • 15 Convenient and validate restriction sites flanking your insert for easy, directional sub-cloning
  • Kanamycin and ampicillin resistance for your choice of selection in E. coli
  • Easy blue⁄white colony screening for selection of recombinants
  • Includes OneShot TOP10 Competent cells for added convenience and highest cloning efficiency
 

pCR2,1- TOPO TA vector.

The pCR2.1-TOPO TA vector

 

Performance and value

Each TOPO TA Cloning Kit contains linearized and topoisomerase I-activated pCR2.1-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 TOPO TA 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, TOPO TA Cloning Kits are available in combo kits combined with a PureLink™ Quick Plasmid Miniprep Kit (50 preps) for fast plasmid purification of your TOPO-cloned inserts for downstream analysis.

Using a proofreading enzyme?

Thermostable polymerases containing extensive 3´ to 5´ exonuclease activity, such as Platinum Pfx, do not leave 3´ A-overhangs. PCR products generated with Taq polymerase have a high efficiency of cloning in the TOPO TA subcloning kits because the 3´ A-overhangs are not removed. However, if you use a proofreading polymerase or wish to clone blunt-ended fragments, you can add 3´ A-overhangs by incubating with Taq at the end of your cycling program.

Alternatively, you may want to try Zero Blunt TOPO PCR Cloning Kits. These kits offer fast and efficient cloning of blunt-end PCR products generated using thermostable, proofreading polymerases. They also utilize the ccdB gene for positive selection.

订购信息

目录号 产品描述 Competent cells supplied 反应次数  
450641
TOPO TA Cloning Kit for Subcloning, without chemically competent cells
20 立即订购
K450001
TOPO TA Cloning Kit for Subcloning, with One Shot TOP10 Chemically Competent E. coli
TOP10 (genotype) 20 立即订购
K452001
TOPO TA Cloning Kit with One Shot MAX Efficiency™ DH5α™ T1 Phage-Resistant Chemically Competent E. coli
MAX Efficiency™ DH5α™ T1R (genotype) 20 立即订购
K455001
TOPO TA Cloning Kit for Subcloning, with One Shot TOP10F' Chemically Competent E. coli TOP10F' (genotype) 20 立即订购
K456001 TOPO TA Cloning Kit for Subcloning, with One Shot TOP10 Electrocomp™ E. coli Electrocompetent TOP10 (genotype) 20 立即订购
K451020
TOPO TA Cloning Kit for Subcloning, with One Shot Mach1™ Phage-Resistant Chemically Competent E. coli Mach1™ T1R (genotype)  20 立即订购
K450002 TOPO TA Cloning Kit with One Shot TOP10 Chemically Competent E. coli and PureLink™ Quick Plasmid Miniprep Kit TOP10 (genotype) 20 立即订购
K451022 TOPO TA Cloning Kit for Subcloning, with One Shot Mach1™ Phage-Resistant Chemically Competent E. coli and PureLink™ Quick Plasmid Miniprep Kit Mach1™ T1R (genotype) 20 立即订购
K450040 TOPO TA Cloning Kit for Subcloning, with One Shot TOP10 Chemically Competent E. coli TOP10 (genotype) 40 立即订购
K452040 TOPO TA Cloning Kit with One Shot MAX Efficiency™ DH5α™ T1 Phage-Resistant Chemically Competent E. coli MAX Efficiency™ DH5α™ T1R (genotype) 40 立即订购
K455040 TOPO TA Cloning Kit for Subcloning, with One Shot TOP10F' Chemically Competent E. coli TOP10F' (genotype) 40 立即订购
K456040 TOPO TA Cloning Kit for Subcloning, with One Shot TOP10 Electrocomp™ E. coli Electrocompetent TOP10 (genotype) 40 立即订购