Support for rapid design of primers for amino acid exchanges and saturation mutagenesis by Golden Gate cloning.
Chris Ulpinnis & Pascal Püllmann
2020-03-05
The Golden Gate cloning technique has been proven to be a highly
efficient toolbox for a variety of cloning setups. Based on its modular
concept it is particularly suitable for the use in multiple-site
mutagenesis approaches. In this technical note we developed a protocol
termed Golden Mutagenesis for the rapid, easy, reliable and cheap
formation of mutagenesis libraries. One to five positions could be
altered in parallel or simultaneously within two days. To facilitate the
implementation of this technique, this R-library has been developed for
the automated primer design and the graphical evaluation of sequencing
results to determine the quality of the library.
The publication is available at https://rdcu.be/bMfta.
A web-based user-interface is available at https://msbi.ipb-halle.de/GoldenMutagenesisWeb.
The GitHub page of the webinterface can be found at https://github.com/ipb-halle/GoldenMutagenesisWeb.
A docker container containing the web interface can be found at https://hub.docker.com/r/sneumann/goldenmutagenesisweb.
You can install GoldenMutagenesis from GitHub with:
# install.packages("devtools")devtools::install_github("ipb-halle/GoldenMutagenesis")
The documentation can be accessed at https://ipb-halle.github.io/GoldenMutagenesis/.
To start with this library we recommend to take a look at our
vignettes!
Point
Mutagenesis
Multiple Site Directed Mutagenesis -
Example1
Multiple Site Directed Mutagenesis -
Example2
Multiple Site Directed Mutagenesis -
Example3
Quick Quality
Control
You can also use predefined Jupiter notebooks on binder to use the library.
