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.

立即订购

 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 pCR®2.1-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