UW Stout/Nystatin SP22

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Pilot Protocol

Purpose: Using a stock solution of nystatin, DMF, and sterile water to treat wild yeast cells. The results of this will be used to decide on a concentration level to apply to different yeast strains.

Materials: 1. Nystatin powder 2. Dimethylformamide (DMF) 3. Sterile water 4. 1.7ml Microcenterfuge tubes 5. Weighing boat 6. Corning COSTAR 96-well clear flat-bottom assay plate 7. Glycerol stock of BY4735 strain as prepared in steps 1-4 of the basic growth curve protocol

Required Equipment: 1. Vortex 2. Scale 3. Molecular Devices SpectraMax Plus 384 Microplate Reader set as outlined in the basic growth curve protocol


Protocol For Pilot Experiment Steps to make stock solution: • weigh out 40mg of Nystatin power • Add nystatin to tube • Measure 10ml DMF • Add DMF to tube • Measure sterile water • Mix until no powder is present


Dilutions for pilot experiment Steps to make dilutions: 1. Mix sterile water with the stock solution, use table below for the amount of each to use 2. Repeat until all concentrations are complete 3. Add 50ul of the diluted product with 50ul of wild yeast

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