Experiment Overview
Repository ID: | FR-FCM-Z3JY | Experiment name: | Expression of pTDH3-YFP in trans-regulatory mutant strains of yeast | MIFlowCyt score: | 24.75% |
Primary researcher: | Fabien Duveau | PI/manager: | Patricia Wittkopp | Uploaded by: | Fabien Duveau |
Experiment dates: | 2017-04-19 - 2017-04-21 | Dataset uploaded: | Mar 2021 | Last updated: | Mar 2021 |
Keywords: | None | Manuscripts: | |||
Organizations: |
University of Michigan, Ann Arbor, MI (USA)
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Purpose: | The goal of this experiment is to compare the expression level of the pTDH3-YFP reporter gene (YFP expressed under control of the TDH3 yeast promoter) between EMS mutants and single-site mutants that only contain a single mutation expected to alter fluorescence from genetic mapping experiments. For each genotype, fluorescence was quantified from 4 replicate populations cultivated in separate wells of 96-well plates at two different dates (8 replicate populations in total). For each replicate population, at least 1000 cells were analyzed by flow cytometry. | ||||
Conclusion: | A strong correlation of pTDH3-YFP expression level is observed between EMS mutants and single-site mutants sharing the same mutation. | ||||
Comments: | None | ||||
Funding: | Not disclosed | ||||
Quality control: | 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. 56 samples correspond to replicate populations of the wild-type progenitor strain used as a reference to quantify the changes of expression 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. |