1 research outputs found
Software-Supported USER Cloning Strategies for Site-Directed Mutagenesis and DNA Assembly
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/