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.