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
- P20, P200 micropipettors
Equipment
Protocol
1. Vortex the yeast culture briefly to resuspend the yeast cells. 2. Set up eight wells according to the following table:
The final concentration of hydroxychloroquine is 300ug/mL. We have controls for this experiment: a well with the wild-type strain yeast untreated with the hydroxychloroquine solution, and wells with the modified strains yeast untreated with the hydroxychloroquine solution for all 8 modified strains. We also have experimental samples: a well with the wild-type strain yeast treated with the hydroxychloroquine solution, and wells with the modified strains yeast treated with the hydroxychloroquine solution for all 8 modified strains. 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.
