Bio-Rad Biofuel Enzyme Kit - Manual (Page 5)

Quick Guide
Activity 3: Determine the effect of
pH on the Reaction Rate
1. Label your cuvettes “pH 5.0”, “pH 6.3”,
and “pH 8.6”.
2. Using a clean DPTP, pipet 500 µl of stop
solution into each cuvette. Wash the
DPTP out thoroughly with water.
3. Using a clean DPTP, pipet 250 µl of
3.0 mM substrate into each microcentrifuge
tube labeled “pH 5.0”, “pH 6.3” and “pH
8.6” by your instructor. Wash the DPTP
out thoroughly with water.
4. Have a stopwatch ready. Using a clean
DPTP, add 250 µl of enzyme to each of
the labeled microcentrifuge tubes.
START YOUR TIMER.
5. After 2 minutes, using a clean DPTP for
each pH reaction, transfer 500 µl of your
reaction to the appropriately labeled
cuvette containing the stop solution.
6. Proceed with the analysis of your samples.
After you have finished your analysis,
rinse out the cuvettes and DPTPs with
copious water and save them for later
activities.
Note: Do not discard unused stock solutions.
They will be used for the next activity.
30
Q
UICK GUIDE
Quick Guide
Label up here
Stop Solution pH 5.0 pH 6.3 pH 8.6
3.0 mM substrate
Enzyme
pH 5.0 pH 6.3 pH 8.6
pH 5.0
pH 5.0
pH 6.3
pH 6.3
pH 8.6
pH 8.6
2 min