Highly efficient, 5-minute cloning with TOPO®

TOPO® TA in vitro transcription kits utlize the pCR®II-TOPO® TA vector with T7 and SP6 promoters for in vitro RNA transcription and sequencing. TOPO® TA technology works by directly ligating Taq-amplified PCR products in a 5-minute, benchtop reaction yielding up to 95% recombinants.

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 The pCR®II-TOPO® TA vector features:

 

  • 3´-T overhangs for direct ligation of Taq-amplified PCR products
  • T7 and SP6 promoters for in vitro RNA transcription and sequencing. The vector also contains M13 forward and reverse primer sites for sequencing.
  • EcoR I sites flanking the PCR product insertion site for easy excision of inserts
  • Kanamycin and ampicillin resistance genes for your choice of selection in E. coli
  • Easy blue/white colony screening for selection of recombinants
 

pCR®-II-TOPO® vector

The pCR®II-TOPO® TA vector

Performance and value

Each TOPO® TA Cloning® for in vitro transcription kit contains linearized and topoisomerase I-activated pCR™II-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.

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  
K460001
TOPO® TA Cloning® Kit for in vitro Transcription, with One Shot® TOP10 Chemically Competent E. coli TOP10 (genotype) 20 立即订购
K462001
TOPO® TA Cloning® Kit for in vitro Transcription, with One Shot® MAX Efficiency™ DH5α T1 Phage-Resistant Chemically Competent E. coli
MAX Efficiency™ DH5α™ T1R (genotype) 20 立即订购
K465001
TOPO® TA Cloning® Kit for in vitro Transcription, with One Shot® TOP10F' Chemically Competent E. coli
TOP10F' (genotype) 20 立即订购
K466001 TOPO® TA Cloning® Kit for in vitro Transcription, with One Shot® TOP10 Electrocomp™ E. coli Electrocompetent TOP10 (genotype) 20 立即订购
K461020
TOPO® TA Cloning® Kit for in vitro Transcription, with One Shot® Mach1™ T1 Phage-Resistant Chemically Competent E. coli
Mach1™ T1R (genotype)  20 立即订购
K460040
TOPO® TA Cloning® Kit for in vitro Transcription, with One Shot® TOP10 Chemically Competent E. coli
TOP10 (genotype) 40 立即订购
K462040 TOPO® TA Cloning® Kit for in vitro Transcription, with One Shot® MAX Efficiency™ DH5α T1 Phage-Resistant Chemically Competent E. coli
MAX Efficiency™ DH5α™ T1R (genotype) 40 立即订购
K465040 TOPO® TA Cloning® Kit for in vitro Transcription, with One Shot® TOP10F' Chemically Competent E. coli
TOP10F' (genotype) 40 立即订购
K466040 TOPO® TA Cloning® Kit for in vitro Transcription, with One Shot® TOP10 Electrocomp™ E. coli
Electrocompetent TOP10 (genotype) 40 立即订购