Difference between revisions of "UW-Stout/Prednisone FA19"
(→Protocol) |
(→Protocol) |
||
Line 28: | Line 28: | ||
#Shake all wells at 37°C for 24 hours. | #Shake all wells at 37°C for 24 hours. | ||
#Plot data and select the Prednisone amount/concentration for the next test that shows results but doesn’t completely kill the yeast cultures. | #Plot data and select the Prednisone amount/concentration for the next test that shows results but doesn’t completely kill the yeast cultures. | ||
+ | |||
+ | ---- | ||
[[File:Dilutions Table.png|800 px]] | [[File:Dilutions Table.png|800 px]] | ||
[[File:Wells Concentrations Table.png|800 px]] | [[File:Wells Concentrations Table.png|800 px]] |
Revision as of 07:28, 12 December 2019
Materials
- Prednisone
- Absolute Ethanol
- Yeast Cultures
- Sterile Water
- Corning COSTAR 96-Well Clear Flat-Bottom Assay Plate
- Micropipettes
- Centrifuge Tubes
Equipment
- Incubator set to 30°C
- Molecular Devices SpectraMax Plus 384 Microplate Reader
Protocol
- Make the stock solution by vortexing the following:
- 50 mg Prednisone
- 10 ml ethanol
- Dilute down to needed percentages (below) using sterile water.
- In each well, add 50 µl of wildtype yeast strain.
- 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.
- 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.
- 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).
- Dilute pure ethanol down to needed percentages.
- In 12 different wells, a new row, add 50 µl of wildtype yeast strain.
- 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.
- Shake all wells at 37°C for 24 hours.
- Plot data and select the Prednisone amount/concentration for the next test that shows results but doesn’t completely kill the yeast cultures.