GENERAL BIOLOGY LAB 1 (BSC1010L)

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a. If you have been assigned Human amylase (H), collect 5mL of saliva from members of your group into a weighing boat. If you ate before coming to lab make sure to rinse out your mouth with water prior to saliva collection. Also, in the event that you are having a hard time producing enough saliva, chew on a small piece of parafilm (it is tasteless). b. Transfer 0.5mL saliva into each test tube. c. Dilute your saliva by adding 0.5mL of distilled water into each test tube containing the saliva. d. If you have been assigned Fungal amylase (F), add 1mL of fungal amylase solution to each tube. 6. Place all 4 test tubes containing starch and the 4 test tubes containing amylase into their respective temperatures. a. 0ºC into the ice bath, b. 40ºC into the 40ºC water bath, c. 60ºC into the 60ºC water bath, d. 95ºC into the 95ºC water bath. 7. Allow all tubes to equilibrate for 5 minutes in their respective temperatures. 8. Add 2-3 drops of iodine to each well at the 0 minutes row. 9. At the end of the equilibration process, without removing the tubes from their water baths, transfer a few drops of the starch solution from each temperature treatment to the first row of the spot plate corresponding to time 0 minutes. Make sure to use a separate transfer pipette for each temperature treatment. Label each of your transfer pipettes with the correct temperature so that they can be reused for each time interval 10. Within each temperature treatment, pour the starch solution into the tube containing amylase. Set your timer for 2 min at the moment of amylase addition. 11. Add 2-3 drops of iodine to each well at the 2 minutes row. This will be repeated before the transfer of each starch-amylase mixture to the spot plates. 12. After 2 min, use the correct transfer pipette for each temperature to remove a few drops of the starch-amylase mixture from each tube. Place 2-3 drops of the mixture in the second row (time = 2 min) on your spot plate under the corresponding temperature. Note the color changes and record your observations in Table 2 (human amylase) or 3 (fungal amylase), depending on your amylase source. Note: You will be analyzing the CLASS DATA SET in your lab reports. 8
13. After each additional 2 min, repeat step 11 and 12. Make sure that you add the starch-amylase mix to the correct wells for time and temperature. 14. At the end of the 10 min, note the temperature and the time at which 100% hydrolysis occurred (Fig. 6). 15. Repeat the procedure using the other amylase type. 16. Use the color-coding scheme below to convert your results (qualitative data) into quantitative (numerical) data. In the column next to your color data, record the corresponding number. STARCH HYDROLYSIS 1 2 3 4 5 MOST STARCH LEAST Figure 6. Starch hydrolysis Table 2: Human amylase Temp ( o C) 0 40 60 95 Color # Color # Color # Color # 0 Time (min) 2 4 6 8 10 9
Page 1 and 2: Exercise 4: Enzymes _______________
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GENERAL BIOLOGY LAB 1 (BSC1010L)

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