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TOPO® Cloning

PCR products can be cloned at high efficiency using TOPO® cloning, which is performed with linearized vectors that have vaccinia virus topoisomerase I covalently bound to the 3’ phosphates. SnapGene simulates three different variations of this method.

TOPO® TA cloning employs linearized vectors such as pcDNA™3.4 TOPO® to clone PCR products with 3’-A overhangs. These PCR products are typically generated with Taq polymerase.

Blunt TOPO® cloning employs linearized vectors such as pCR®-Blunt II-TOPO® to clone PCR products with blunt overhangs. These PCR products are typically generated with high-fidelity polymerases.

Directional TOPO® cloning employs linearized vectors such as pET200/D-TOPO® to clone PCR products in a directional manner.

SnapGene simplifies TOPO® cloning by automating the primer design. To plan a TOPO® cloning procedure, select the DNA fragment that you wish to amplify, and choose a TOPO® cloning vector from a list that is embedded in the software. SnapGene will then choose suitable primers.