Quick and efficient PCR cloning with TOPO® technology

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
  • EcoRI sites flanking the PCR product insertion site for easy excision of inserts
  • 15 convenient and validated 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 diagram

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.

Ordering information

Cat. No. Product description Competent cells supplied Number of reactions  
451641 TOPO® TA Cloning® Kit for Subcloning None 10 Order now
K4500J10 TOPO® TA Cloning® Kit for Subcloning TOP10 (genotype) 10 Order now
450641
TOPO® TA Cloning Kit for Subcloning, without chemically competent cells
None 25 Order now
K450001
TOPO® TA Cloning Kit for Subcloning, with One Shot® TOP10 Chemically Competent E. coli
TOP10 (genotype) 25 Order now
K452001
TOPO® TA Cloning Kit with One Shot® MAX Efficiency™ DH5α™ T1 Phage-Resistant Chemically Competent E. coli
MAX Efficiency™ DH5α™ T1R (genotype) 25 Order now
K455001
TOPO® TA Cloning Kit for Subcloning, with One Shot® TOP10F' Chemically Competent E. coli TOP10F' (genotype) 25 Order now
K456001 TOPO® TA Cloning Kit for Subcloning, with One Shot® TOP10 Electrocomp™ E. coli Electrocompetent TOP10 (genotype) 25 Order now
K451020
TOPO® TA Cloning Kit for Subcloning, with One Shot® Mach1™ Phage-Resistant Chemically Competent E. coli Mach1™ T1R (genotype)  25 Order now
K450002 TOPO® TA Cloning® Kit with One Shot® TOP10 Chemically Competent E. coli and PureLink™ Quick Plasmid Miniprep Kit TOP10 (genotype) 25 Order now
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) 25 Order now
K450040 TOPO® TA Cloning Kit for Subcloning, with One Shot® TOP10 Chemically Competent E. coli TOP10 (genotype) 50 Order now
K452040 TOPO® TA Cloning Kit with One Shot® MAX Efficiency™ DH5α™ T1 Phage-Resistant Chemically Competent E. coli MAX Efficiency™ DH5α™ T1R (genotype) 50 Order now
K455040 TOPO® TA Cloning Kit for Subcloning, with One Shot® TOP10F' Chemically Competent E. coli TOP10F' (genotype) 50 Order Now
K456040 TOPO® TA Cloning Kit for Subcloning, with One Shot® TOP10 Electrocomp™ E. coli Electrocompetent TOP10 (genotype) 50 Order now