Haploids were obtained by sporulation, dissection onto YPDA, replica plated onto complete supplement media minus tryptophan, and tryptophan prototroph colonies selected.
The strains in each plate were spotted onto solid YPD medium, then replica printed onto YPD medium with or without 0.1 mg/mL hygromycin B for the primary screen.
At least 14 individual colonies for each construct were plated onto -Ura master plate which was replica plated onto YPD, YPG, and –Ade plates to monitor color pigmentation, petite phenotype and growth on -Ade, respectively.
The cells were then rehydrated by placing the filter onto a YPD plate, the cells were replica plated onto a second YPD plate and survival assessed by growth on the replica plate.
The colony patches were replica plated onto NCE-glucose medium to assess the frequency of auxotrophs and onto MacConkey-fructose, MacConkey-maltose, MacConkey-propanediol, and MacConkey-xylose agar plates to identify fermentation mutations.
After different induction periods, cells were plated onto selective media and [PSI+] phenotypes were scored after replica plating onto rich media as well as media lacking adenine.
After replica plating onto SG HKL medium, colonies presenting a growth defect were isolated and spotted individually.
Established colonies were replica plated onto low potassium plates and challenged with different concentrations of blocker applied via Whatman filters.
After 72 h growth, colonies were replica plated onto plates containing GM17 with erythromycin, tetracycline or chloramphenicol.
The resulting colonies were replica plated onto the selective medium to determine the relative percent of each strain recovered.
