Yeast transformation
For selection of transformants of the Cas9 plasmid, and maintenance in
SC-Rep. Transformation is done according to a modified one-step lithium
acetate transformation protocol from (Gietz et al. , 1995). The
duration of the heat shock at 42°C differs according to the species; 25
min for C. glabrata , 20 min for C. bracarensis , 15 min forC. nivariensis .
For induction of DSBs, all species were transformed with the pCFYF
plasmid into which is cloned the gRNA specific for each species, but
also with the plasmid that does not express any gRNA, to control for
medium/plasmid/Cas9 expression effects (see Results).