Software-Supported USER Cloning Strategies for Site-Directed
Mutagenesis and DNA Assembly
Version 3 2018-02-10, 00:03
Version 2 2017-10-17, 18:33
Version 1 2016-02-14, 13:37
Posted on 2018-02-10 - 00:03
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/.
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Genee, Hans Jasper; Bonde, Mads Tvillinggaard; Bagger, Frederik
Otzen; Jespersen, Jakob
Berg; O. A. Sommer, Morten; Wernersson, Rasmus; et al. (2016). Software-Supported USER Cloning Strategies for Site-Directed
Mutagenesis and DNA Assembly. ACS Publications. Collection. https://doi.org/10.1021/sb500194z
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AUTHORS (7)
HG
Hans Jasper Genee
MB
Mads Tvillinggaard Bonde
FB
Frederik
Otzen Bagger
JJ
Jakob
Berg Jespersen
MO
Morten O. A. Sommer
RW
Rasmus Wernersson
LO
Lars Rønn Olsen