UW-Stout/Caffeine SP21
Contents
Materials
- Caffeine 80 mM solution in water. (15.52 x 25 ml water= .388 g of caffeine)
- Sterile H2O
- Wild-type yeast in 2x synthetic complete media at an OD600 of 0.1-0.2
- Corning COSTAR 96-well clear flat-bottom assay plate
- 250 mL beaker for caffeine master solution.
- P-10,200 micro pipette/ tips
- Scale
- Tube to store caffeine
- Powdered caffeine for solution
Equiptment
- Molecular Devices SpectraMax Plus 384 Microplate Reader
Protocol
- Weigh out .388 grams of caffeine
- Add to 25 ml of water in test tube and swirl until it’s mixed with no chunks
- Set up 8 test tubes and label them 1 through 8 matching with the order of wells
- Using the p20 pipette, pipette the varying amounts of caffeine solution provided in table
- Then using a p200 pipette, pipette the varying amount of sterile water into the provided tubes
- When under the hood add 50 ul of yeast to each test tube except for the tube labeled 0 which is the control
- Transfer the solution created in each tube into the wells, writing down which row your wells are
- Set up the plate reader as follows:
Temperature: 30°C Mode: Kinetic Wavelength: 600 nm Interval: 10 minutes Total run time: 12 hours Shake before read: 30 seconds Transfer the assay plate to the reader and read for 24 hours. This process will be replicated 3 times.
Data
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