Difference between revisions of "UW-Stout/Caffeine SP21"
Line 26: | Line 26: | ||
#Transfer the solution created in each tube into the wells, writing down which row your wells are | #Transfer the solution created in each tube into the wells, writing down which row your wells are | ||
#Set up the plate reader as follows: | #Set up the plate reader as follows: | ||
− | Temperature: 30°C | + | *Temperature: 30°C |
− | Mode: Kinetic | + | *Mode: Kinetic |
− | Wavelength: 600 nm | + | *Wavelength: 600 nm |
− | Interval: 10 minutes | + | *Interval: 10 minutes |
− | Total run time: 12 hours | + | *Total run time: 12 hours |
− | Shake before read: 30 seconds | + | *Shake before read: 30 seconds |
− | Transfer the assay plate to the reader and read for 24 hours. | + | *Transfer the assay plate to the reader and read for 24 hours. |
This process will be replicated 3 times. | This process will be replicated 3 times. | ||
== Data == | == Data == |
Revision as of 10:44, 20 April 2021
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.