Team:Calgary/Notebook/Protocols/PrussianBlueFerritinTemperatureOptimization
From 2013.igem.org
(Difference between revisions)
Line 21: | Line 21: | ||
<li>Add the appropriate amount of hydrogen peroxide, and IMMEDIATELY begin taking readings.</li> | <li>Add the appropriate amount of hydrogen peroxide, and IMMEDIATELY begin taking readings.</li> | ||
<li>Repeat the experiment multiple times for each temperature (25-45˚C with 5˚C intervals)</li> | <li>Repeat the experiment multiple times for each temperature (25-45˚C with 5˚C intervals)</li> | ||
- | <li>Slopes of each experiment was determined and plotted | + | <li>Slopes of each experiment was determined and plotted against temperature to determine optimum temperature.</li> |
</ol> | </ol> | ||
Latest revision as of 00:48, 28 September 2013
Temperature Optimization
Prussian Blue Ferritin Temperature Optimization
Reaction Mixture
- 10μL Prussian Blue Ferritin (22μg/mL)
- 6μL Substrate (TMB or ABTS, 10mg/mL)
- 32μL Hydrogen Peroxide (30%)
- Sodium Acetate-Acetic Acid Buffer (pH 3.6) up to 242μL
- Combine all of the reagents listed in the reaction mixture, minus hydrogen peroxide.
- Set the spectrophotomer to read the appropriate absorbance (650nm for TMB, 415nm for ABTS) for 10 minutes with 10 second intervals.
- Add the appropriate amount of hydrogen peroxide, and IMMEDIATELY begin taking readings.
- Repeat the experiment multiple times for each temperature (25-45˚C with 5˚C intervals)
- Slopes of each experiment was determined and plotted against temperature to determine optimum temperature.