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Region Hovedstaden - en del af Københavns Universitetshospital
Udgivet

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

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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 (Automated DNA Modifications with USER 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/ .

OriginalsprogEngelsk
TidsskriftA C S Synthetic Biology
Vol/bind4
Udgave nummer3
Sider (fra-til)342-9
Antal sider8
ISSN2161-5063
DOI
StatusUdgivet - 20 mar. 2015

ID: 45103457