Design Genes with Ease using In-Fusion® Cloning by Clontech Laboratories, a Takara Bio Company
23 July 2015

In-Fusion can join any two pieces of DNA that contain a 15 bp overlap at their ends. The result is equivalent to a recombination event across the 15 bp overlap. The 15 bp overlap can be engineered by inclusion in forward and reverse primers used to PCR-amplify two segments of DNA. Originally described for inserting one piece of DNA into a restriction enzyme-digested plasmid, it has been found that In-Fusion can efficiently join up to four pieces of DNA in a single reaction. This application note explores the unique end-joining capabilities of In-Fusion to combine four fragments of DNA in a single, one-step reaction to develop seamless fusion proteins and modular expression vectors.