UW-Stout/Caffeine FA19
Caffeine Dilution and Concentration of Caffeine Protocol
Materials
- 0.22g Caffeine
- Sterile H2O
- Syringes and purifier
- 96 Well assay plate
Procedure
- Make a 100μM stock solution of caffeine and H2O.
- Take 0.1942 grams of caffeine to 10ml of H2O to form a stock solution of 100C.
- Make another stock solution of 10μM of caffeine and H2O.
- Dilute the 100μM stock solution of 10μM stock solution by adding 4μl of 100μM of the stock solution and 36μl sterile H20.
- Filter the caffeine solution using the syringes and purifier then put in microfuge tubes, freeze at -20°C.
- Dilute the 100μM stock solution into the 12 concentrations using the table below.
- Fill the wells with different concentrations of caffeine solution.
- After the wells are completed, place the 96 Well assay plate into the SpectraMax Plus 384 Microplate Reader. Data will be collected every 5 minutes for the next 24 hours.
- Once results are computed the data curve should be put on the growth data curve
Volume of the Stock solution of 10mM (microliters) 0 50 50 - 0 0.10 50 49 - 1 0.25 50 47.5 - 2.5 0.50 50 45 - 5 1.0 50 40 - 10 2.5 50 47.5 2.5 - 5.0 50 45 5 - 8.0 50 42 8 - 10 50 40 10 - 15 50 35 15 - 20 50 30 20 - 50 50 0 50 -