To begin the experimental set up, grab a napkin/paper and place it under the spot plates. Across the top of the spot plates, on the napkin, write Temperature and label each column using 0°, 25°, 55°, and 85°. Along side the spot plates write Time and label each row using 0, 2, 4, 6, 8, and 10 minutes. Next, acquire four different test tubes and label each of them with a different temperature, label the enzyme source (B for bacterial or F for fungal), add the group number. Obtain another 4 test tubes and do the same as the previous step but add the letter S to signify starch solution. For the test tubes labeled S, add 5 mL of starch solution. To continue to the effect of temperature on amylase activity, add 1 mL of amylase into each of the
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If assigned to test the bacterial amylase first, place the amylase solution in the bacterial amylase designated test tubes. If assigned to do fungal amylase first, place the amylase solution into the fungal amylase designated test tubes. Place all of the test tubes, no matter whether they carry starch of amylase, and place them into their respective temperatures depending on how they are labeled. Allow all of the tubes to equilibrate in their temperature baths for 5 minutes. During these 5 minutes, add 3 drops of iodine into each well at the 0 minutes row, the first well of each temperature column. Once the 5 minutes are up, without removing the tubes from their temperature baths, use a pipette to transfer 3 drops of the starch solution from each temperature treatment to the 0 minutes row where the iodine was recently placed. Label each of the transfer pipettes with the corresponding temperature so they can be reused for each time interval but make sure to use a separate pipette for each temperature treatment. During the temperature treatment, pour the starch solution into the containing amylase and set the timer for 2 minutes. Add 3 drops of iodine to each well at the 2 minutes row. This will be done before the transfer of each
If assigned to test the bacterial amylase first, place the amylase solution in the bacterial amylase designated test tubes. If assigned to do fungal amylase first, place the amylase solution into the fungal amylase designated test tubes. Place all of the test tubes, no matter whether they carry starch of amylase, and place them into their respective temperatures depending on how they are labeled. Allow all of the tubes to equilibrate in their temperature baths for 5 minutes. During these 5 minutes, add 3 drops of iodine into each well at the 0 minutes row, the first well of each temperature column. Once the 5 minutes are up, without removing the tubes from their temperature baths, use a pipette to transfer 3 drops of the starch solution from each temperature treatment to the 0 minutes row where the iodine was recently placed. Label each of the transfer pipettes with the corresponding temperature so they can be reused for each time interval but make sure to use a separate pipette for each temperature treatment. During the temperature treatment, pour the starch solution into the containing amylase and set the timer for 2 minutes. Add 3 drops of iodine to each well at the 2 minutes row. This will be done before the transfer of each