TOPO TA for Subcloning

Quick and efficient PCR cloning with TOPO technology

TOPO TA subcloning kits utilize our popular pCR 2.1-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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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. TheTOPO 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.
 

Thermostable polymerases containing extensive 3´ to 5´ exonuclease activity, such as Platinum SuperFi, 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.

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