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!
Multiple Site Directed Mutagenesis - Example1
Multiple Site Directed Mutagenesis - Example2
Multiple Site Directed Mutagenesis - Example3
Quick Quality Control