Simulate TOPO Cloning

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.

Directional TOPO® cloning

Simulate TOPO Cloning in SnapGene

Learn More

► Simulate TA and GC Cloning

Learn how to simulate TA and GC cloning in SnapGene

► Simulating Cloning in SnapGene

A video series on how to simulate a range of cloning techniques in SnapGene

► SnapGene User Guides

Step by step instructions for TOPO cloning with SnapGene

► TOPO Cloning guide

An in-depth guide to performing TOPO cloning

► Polymerase Chain Reaction Guide

An in-depth guide to the polymerase chain reaction (PCR)