TY - JOUR
T1 - Software-Supported USER Cloning Strategies for Site-Directed Mutagenesis and DNA Assembly
AU - Genee, Hans Jasper
AU - Bonde, Mads Tvillinggaard
AU - Bagger, Frederik Otzen
AU - Jespersen, Jakob Berg
AU - Sommer, Morten
AU - Wernersson, Rasmus
AU - Olsen, Lars Rønn
PY - 2015/3/20
Y1 - 2015/3/20
N2 - 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/ .
AB - 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/ .
U2 - 10.1021/sb500194z
DO - 10.1021/sb500194z
M3 - Journal article
C2 - 24847672
SN - 2161-5063
VL - 4
SP - 342
EP - 349
JO - A C S Synthetic Biology
JF - A C S Synthetic Biology
IS - 3
ER -