Difference between revisions of "UW-Stout/Hydroxychloroquine SP21"
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# Vortex the yeast culture briefly to resuspend the yeast cells. | # Vortex the yeast culture briefly to resuspend the yeast cells. | ||
# 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. | # 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. |
#To measure cell growth over a period of 24 hours, set up the plate reader as follows: | #To measure cell growth over a period of 24 hours, set up the plate reader as follows: | ||
#*Temperature: 30°C | #*Temperature: 30°C | ||
Revision as of 08:07, 27 April 2021
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
- Vortex the yeast culture briefly to resuspend the yeast cells.
- 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.
- 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
- Transfer the assay plate to the reader and read for 24 hours.
- Note: Repeat the experiment a few times to account for natural variation.
