Software-Supported USER Cloning Strategies for Site-Directed Mutagenesis and DNA Assembly

Published on 2018-02-10T00:03:29Z (GMT) by
USER cloning is a fast and versatile method for engineering of plasmid DNA. We have developed a user friendly Web server tool that automates the design of optimal PCR primers for several distinct USER cloning-based applications. Our Web server, named AMUSER (<u>A</u>utomated DNA <u>M</u>odifications with <u>USER</u> cloning), facilitates DNA assembly and introduction of virtually any type of site-directed mutagenesis by designing optimal PCR primers for the desired genetic changes. To demonstrate the utility, we designed primers for a simultaneous two-position site-directed mutagenesis of green fluorescent protein (GFP) to yellow fluorescent protein (YFP), which in a single step reaction resulted in a 94% cloning efficiency. AMUSER also supports degenerate nucleotide primers, single insert combinatorial assembly, and flexible parameters for PCR amplification. AMUSER is freely available online at http://www.cbs.dtu.dk/services/AMUSER/.

Cite this collection

Genee, Hans Jasper; Bonde, Mads Tvillinggaard; Bagger, Frederik

Otzen; Jespersen, Jakob

Berg; O. A. Sommer, Morten; Wernersson, Rasmus; Olsen, Lars Rønn (2018): Software-Supported USER Cloning Strategies for Site-Directed

Mutagenesis and DNA Assembly. ACS Publications. Collection.