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
-