Bio-Rad Biofuel Enzyme Kit - Manual (Page 8)

Quick Guide
Activity 5: Determine the Effect of
Substrate Concentration on the
Reaction Rate
1. Label one clean 15 ml conical tube “Low
Concentration Substrate” and one clean
15 ml conical tube “High Concentration
Substrate”.
2. Using a clean DPTP, pipet 1.5 ml of 1.5 mM
substrate into the 15 ml conical tube
labeled “High Concentration Substrate”.
Rinse the DPTP thoroughly with clean
water.
3. Using a clean DPTP, pipet 1.25 ml of
buffer into the 15 ml conical tube labeled
“Low Concentration Substrate”. Rinse
the DPTP thoroughly with water and then
pipet 250 µl of 1.5 mM substrate into the
15 ml conical tube labeled “Low
Concentration Substrate” and mix. Rinse
the DPTP thoroughly with water.
4. Label your cuvettes “H1–H3” (for high
substrate concentration time points) and
“L1–L3” (for low substrate concentration
time points). Only label on the upper part
of the cuvette face.
5. Using a clean DPTP, pipet 500 µl of stop
solution into each cuvette. Rinse the
DPTP thoroughly with water.
6. Label one DPTP as “H” for high substrate
concentration and a second DPTP as “L”
for low substrate concentration.
33
Q
UICK GUIDE
Quick Guide
1.5 mM Substrate High concentration substrate
Buffer 1.5 mM Substrate Low
concentration
substrate
Label up here
Stop Solution H1 H2 H3 L1 L2 L3