Chris Ulpinnis & Pascal Püllmann 2020-03-05

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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


A web-based user-interface is available at

The GitHub page of the webinterface can be found at

A docker container containing the web interface can be found at


You can install GoldenMutagenesis from GitHub with:

# install.packages("devtools")


The documentation can be accessed at


Jupiter Notebooks at binder

You can also use predefined Jupiter notebooks on binder to use the library.

Multiple Site Directed Mutagenesis


Single Point Mutagenesis


Domestication only