Secondary screen of pTDH3-YFP expression changes in yeast EMS mutants

The purpose of the experiment was to identify EMS mutants strains with altered expression of the pTDH3-YFP reporter gene (YFP expressed under control of the TDH3 promoter in yeast cells). 6 replicate populations of 315 mutant strains were grown separately in YPD medium before measuring fluorescence of >1000 cells for each sample on a BD C6 Accuri instrument.
Conclusion:
Quantitative analysis of flow cytometry data showed a significant change of expression in 174 of the 315 mutants. 44 of these mutants were then selected for mapping mutations altering pTDH3-YFP expression in the yeast genome. The experiment includes several replicate populations of a strain that does not contain the pTDH3-YFP reporter gene and that is used to estimate the level of autofluorescence. 120 samples correspond to replicate populations of the wild-type progenitor strain used as a reference to quantify expression changes in each mutant. An additional 24 wild-type strains expressing pTDH3-YFP were arrayed at predetermined positions in each 96-well plates to correct for technical variation between plates or flow cytometry runs.