Bio-Rad Biofuel Enzyme Kit - Manual (Page 11)

7. Using a clean DPTP, pipet 500 µl of stop
solution into each cuvette. Rinse out the
DPTP thoroughly with water.
8. Label a 15 ml conical tube with the type
of mushroom you are using and then
using a clean DPTP, pipet 3 ml of substrate
into the tube.
Please read and understand steps 9–10
fully before proceeding. These steps are
time sensitive!
9. Using a clean DPTP, pipet 250 µl of your
enzyme extract into your 15 ml conical
tube of substrate. START YOUR TIMER.
10. At the times indicated, remove 500 µl of
mushroom extract/substrate mixture from
the reaction tube, and add it to the
appropriately labeled cuvette that already
contains the stop solution.
11. Using a clean DPTP, add 500 µl of
extraction buffer to cuvette #6. Clean the
DPTP and then add one drop of mushroom
extract. This will serve as the “blank” for
this experiment.
12. Proceed with the analysis of your
samples. After you have finished your
analysis, rinse out the reaction tubes,
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.
36
Q
UICK GUIDE
Quick Guide
Stop Solution 1 2 3 4 5 6
Reaction
Tube
1
1 min
2
2 min
3
4 min
4
6 min
5
8 min
1.5 mM Substrate Empty reaction tube
Mushroom extract Reaction tube
with substrate
Extraction
buffer
Cuvette #6 Mushroom extract