Site-Directed Mutagenesis: PCR Based Mutagenesis with the In-Fusion HD Cloning System
In-Fusion Cloning Kits enable PCR mediated site-directed DNA mutagenesis. You can add, delete, or replace DNA segments; insert point mutations; insert restriction sites and more using a streamlined single reaction protocol.
The In-Fusion method relies on the creation of primers that incorporate the mutation (base pair changes, deletions, additions) and a gene-specific priming sequence. Using an inverse PCR reaction with a circular plasmid as the template, a linearized vector is created that incorporates the mutation. The linearized vector is re-circularized via the In-Fusion reaction. High-efficiency competent cells are also provided with the kits.