UW-Stout/Prednisone FA19

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Revision as of 08:33, 12 December 2019 by Teagueb (talk | contribs) (Analysis)
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  • Prednisone
  • Absolute Ethanol
  • Yeast Cultures
  • Sterile Water
  • Corning COSTAR 96-Well Clear Flat-Bottom Assay Plate
  • Micropipettes
  • Centrifuge Tubes


  • Incubator set to 30°C
  • Molecular Devices SpectraMax Plus 384 Microplate Reader


  1. Make the stock solution by vortexing the following:
    • 50 mg Prednisone
    • 10 ml ethanol
  2. Dilute down to needed percentages (below) using sterile water.
  3. In each well, add 50 µl of wildtype yeast strain.
  4. Using the stock solution (P1), add 20 µl to the first well along with 30 µl of sterile water.
    • This well should now have 100 µl total of solution.
  5. Using the stock solution (P1), add 6 µl to the second well along with 44 µl of sterile water.
    • This well should now have 100 µl total of solution.
  6. Repeat this process for each set of wells, going down one dilution after each set of two wells.
    • Refer to the Well Concentrations table (below).
  7. Dilute pure ethanol down to needed percentages.
  8. In 12 different wells, a new row, add 50 µl of wildtype yeast strain.
  9. Add the ethanol dilutions to the 12 new wells, using the same process as used above.
    • This row will be used to compare the Prednisone treated yeasts’ growth rates.
  10. Shake all wells at 37°C for 24 hours.
  11. Plot data and select the Prednisone amount/concentration for the next test that shows results but doesn’t completely kill the yeast cultures.

Dilutions Table.png

Wells Concentrations Table.png


Prednisone Graph.png Ethanol Graph.png


Using the ethanol tests as a comparison to evaluate the effects of Prednisone, it is shown that the Prednisone had a substantial effect on the growth of the wild-type yeast strain (BY4735). Many of the higher concentrations of Prednisone negatively affected the yeast, inhibiting the growth of the strain, while the lower concentrations of Prednisone actually increased the growth concentrations. Wells with the largest concentrations of ethanol experienced no growth in both graphs, showing that the ethanol was the greatest influence on the strain. The third-largest concentration (100.0 µg/ml) also inhibited the growth, but the concentrations were very similar throughout the tests. On the other hand, smaller concentrations such as the well with 0.3 µg/ml increased the growth of BY4735 compared to the effects of only ethanol at this concentration. These results were quite surprising, as we expected the Prednisone to inhibit the growth rates more than ethanol alone.