M465:IndependentProject

For those unfamiliar with microbiology

The Effect of Environment on the Microbiota

For your independent project, you have chosen to fund a proposal focused on environmental variables that may alter or impact the Drosophila microbiota. Each team will focus on one of the three variables below:
1) Temperature (18C, 25C, 28C)
2) Glyphosate (0 ug/mL, 0.1 ug/mL, 0.5 ug/mL)
3) Antibiotics (0 ug/mL, 5 ug/mL, 50 ug/mL)

For each variable, you will have three conditions (outlined above in parentheses). You will begin with wild type flies lacking any Wolbachia infection.

Below is a rough outline of the next few weeks - things may be adjusted depending on how well the flies grow, etc.

Tuesday, October 25th: Set up flies

Today we'll be establishing our flies in each of our conditions. Below are protocols for each of the three variables above. Remember that each team will focus on one of these variables. Each student will have their own set of three experimental lines for this experiment.

Glyphosate

• Protocol to be performed as a group such that you standardize your food across the team.
  

1. Each student will collect three vials of food from your instructor. These vials have 5 mL of food in them each. In addition, you will need two small beakers.
2. Microwave the food vials for 10 seconds at a time until the food is liquified. Pour the food into the small beaker.
3. Using a plastic graduated pipette, take 5 mL of the food and place into one of the previously used glass vials. Label this vial CONTROL. Create four control vials
4. Calculate the amount of glyphosate needed for the remaining food. For example, if you have four people in your group, you will have liquified total of 60 mL of food, out of which you will have used 20 mL for the CONTROL vials. That would leave 40 mL. Remove 20 mL of food and place in a new beaker.
5. Create a food mixture with 0.1 ug/mL glyphosate and one with 0.5 ug/mL glyphosate. Stir well and dispense using the graduated pipettes.
6. Each student in the group should have one control, one 0.1 ug/mL vial and one 0.5 ug/mL vial.

Collecting your flies

Make sure your vials are well labeled with your name, fly stock, and germ-free and put them on the front bench. We'll take them to the Newton lab to hatch.