Team:Montpellier/Protocols
PROTOCOLS
In order to perform our proof of concept, we first had to assemble and clone all the constructs in the chosen vectors. We did this by Golden gate and Gibson assembly, first in chemically competent DH5-Alpha E. coli and later in electrocompetent NEB10-Beta E. coli in order to increase the transformation efficiency. Next, we induced the expression of the proteins of interest for each plasmid and, after cotransformations of the reporters with KARMA or TEV, we measured the corresponding fluorescence of GFP or RFP with a plate reader.
Here we present the detailed protocols used for each technique, as well as for the routine tasks we performed in the lab. Most of those protocols are adapted from the Bonnet Team Protocol Repository.
Summary:
