Bio-Rad Biofuel Enzyme Kit - Manual (Page 3)

Quick Guide
Activity 2: Determine the Effect of
Temperature on the Reaction Rate
1. Label your cuvettes “0°C”, “22°C”, and
“37°C”.
2. Using a clean DPTP, pipet 500 µl of stop
solution into each cuvette. Wash the
DPTP out thoroughly with water.
3. Label three 1.5 ml microcentrifuge tubes
“0°C Enzyme”, “22°C Enzyme”, and
“37°C Enzyme”. Using a clean DPTP,
pipet 250 µl of enzyme into each
microcentrifuge tube. Rinse out the
DPTP thoroughly with water.
4. Label three 1.5 ml microcentrifuge tubes
“0°C Substrate”, “22°C Substrate”, and
“37°C Substrate”. Using a clean DPTP,
pipet 500 µl of 1.5 mM substrate into
each microcentrifuge tube. Rinse out the
DPTP thoroughly with water.
5. Place the tubes labeled “0°C Enzyme”
and “0°C Substrate” in the ice cup. Place
the tubes labeled “22°C Enzyme” and
“22°C Substrate” on your lab bench.
Place the tubes labeled “37°C Enzyme”
and “37°C Substrate” in the beaker of
warm water at 37°C. Allow the tubes to
equilibrate to their respective tempera-
tures for at least 5 minutes.
28
Q
UICK GUIDE
Quick Guide
Label up here
Stop Solution 0°C 22°C 37°C
Enzyme 0°C
Enzyme
22°C
Enzyme
37°C
Enzyme
1.5 mM Substrate 0°C
Substrate
0°C
Substrate
22°C
Substrate
37°C
Substrate
0°C
Enzyme
0°C
22°C
Enzyme
Room
Temperature
37°C
Enzyme
37°C
22°C
Substrate
37°C
Substrate