Bio-rad Biofuel Enzyme Kit Manual de usuario descargar pdf (Pagina 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

QUICK 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

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