Bio-rad Biofuel Enzyme Kit Manuel d'utilisateur télécharger pdf (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

QUICK 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

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