UW-Stout/Hydroxychloroquine SP21

Materials

• Corning COSTAR 96-well clear flat-bottom assay plate
• Wild-type yeast in 2x synthetic complete media at an OD600 of 0.1-0.2
• Hydroxychloroquine, 300 ug/uL solution in sterile water (as calculated in the calibration protocol)
• P20, P200 micropipettors

Equipment

• Molecular Devices SpectraMax Plus 384 Microplate Reader

Precautions

Use gloves and safety glasses for this experiment. Hydroxychloroquine is a potent chemical used to treat malaria and some autoimmune conditions. However, side effects include but are not limited to nausea, headache, dizziness, rash.

Protocol

1. Vortex the yeast culture briefly to resuspend the yeast cells.
2. Set up seven wells aliquoting 50uL of the stock hydroxychloroquine and 50uL of yeast culture. Use the seventh well for the wild-type yeast strain.
   • The final concentration of hydroxychloroquine is 300ug/mL.
3. To measure cell growth over a period of 24 hours, set up the plate reader as follows:
   • Temperature: 30°C
   • Mode: Kinetic
   • Wavelength: 600 nm
   • Interval: 5 minutes
   • Total run time: 24 hours
   • Shake before reading: 30 seconds
4. Transfer the assay plate to the reader and read for 24 hours.
   • Note: Repeat the experiment a few times to account for natural variation.