Bio-Rad Biofuel Enzyme Kit - Manual (Page 4)

6. Have a stopwatch ready. Using a clean
DPTP, pipet the 250 µl of enzyme from
the tube labeled “0°C Enzyme” into the
tube labeled “0°C Substrate”, and place
the tube now containing your enzyme
and substrate mix back on ice. Add the
22°C enzyme to the 22°C substrate
solution, and place that tube back on the
lab bench. Add the 37°C substrate to the
37°C enzyme solutions, and put that tube
back into the 37°C water bath. START
YOUR TIMER.
7. After 2 minutes, use a clean DPTP for
each temperature reaction to transfer
500 µl of your reaction to the appropriately
labeled cuvette containing the stop
solution.
8. 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.
29
Quick Guide
Q
UICK GUIDE
0°C
0°C S 22°C S 37°C S 0°C E
0°C
22°C E
Room
Temperature
37°C E
37°C
0°C
S+E
22°C 22°C
S+E
37°C 37°C
S+E
2 min