Effect of pH on the Rate of Reaction of Amylase (Required Practical)
1. Use the marker pen to label a test tube with the first value of pH buffer solution (pH 4) and stand it in the test tube rack
2. Into each well of the spotting tiles, place a drop of iodine
3. Using a measuring cylinder, measure 2cm of amylase and pour into the test tube
4. Using a syringe, measure 1cm' of the buffer solution and pour into the test tube
5. Leave this to stand for five minutes and then use the thermometer to measure the temperature. Make a note of the temperature
6. Add 2cm' of starch solution into the test tube, using a different measuring cylinder to measure, and begin a timer (leave the timer to run continuously)
7. After 10 seconds, use a pipette to extract some of the amylase/starch solution and place one drop into the first well of the spotting tile. Squirt the remaining solution back into the test tube
8. Continue to place one drop into the next well of the spotting tile, every 10 seconds, until the iodine remains orange
9. Record the time taken for the starch to be completely digested by the amylase by counting the wells that were tested positive for starch (indicated by the blue/black colour change of the iodine). Fach well represents 10 seconds of time
10. 10. Repeat steps 1 to 8 for pH values 7 and 10