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 .
Original languageEnglish
JournalA C S Synthetic Biology
Volume4
Issue number3
Pages (from-to)342-349
Number of pages8
ISSN2161-5063
DOIs
Publication statusPublished - 2015
CitationsWeb of Science® Times Cited: No match on DOI

    Research areas

  • DNA assembly, USER cloning, Primer design, Site-directed mutagenesis, Point mutation, Web server

ID: 92848953