Difference between revisions of "UW-Stout/Hydroxychloroquine SP21"

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(→‎Protocol)
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==Protocol==
 
==Protocol==
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1. Vortex the yeast culture briefly to resuspend the yeast cells.
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# Vortex the yeast culture briefly to resuspend the yeast cells.
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2. Set up eight wells according to the following table:
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# 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.
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##The final concentration of hydroxychloroquine is 300ug/mL.  
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[[File:Capture.png]]
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#To measure cell growth over a period of 24 hours, set up the plate reader as follows:
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#*Temperature: 30°C
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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.
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#*Mode: Kinetic
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3. To measure cell growth over a period of 24 hours, set up the plate reader as follows:
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#*Wavelength: 600 nm
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Temperature: 30°C
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#*Interval: 5 minutes
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Mode: Kinetic
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#*Total run time: 24 hours
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Wavelength: 600 nm
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#*Shake before reading: 30 seconds
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Interval: 5 minutes
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#Transfer the assay plate to the reader and read for 24 hours.
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Total run time: 24 hours
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#*Note: Repeat the experiment a few times to account for natural variation.
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Shake before reading: 30 seconds
 
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4. Transfer the assay plate to the reader and read for 24 hours.
 
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Note: Repeat the experiment a few times to account for natural variation.
 

Revision as of 08:07, 27 April 2021

Materials

Equipment

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.
    1. 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.
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