Larvae Collection and Prelabeling Treatment

1. Drosophila media (Carolina Biological Supply Co.): You will need bags of both the blue and white instant foods. Cultures for egg-laying are done in plastic bottles (Applied Scientific) that can fit a 35-mm culture dish on top.

2. Grape plates for egg-laying: 100 mL Welch's grape juice, 100 mL bi-distilled water (ddH2O), 8 g agar. Melt in a microwave, cool to approx 60°C, and add 1 mL acetic acid and 1 mL 100% ethanol. Mix and pour into 35-mm culture dishes. Store in plastic boxes at 4°C for up to 5 d. Bring plates to room temperature before use.

3. Sucrose food: 20% Sucrose in phosphate-buffered saline (PBS), 1.5% agar. Aliquot approx 10 mL into small plastic vials. Plug with cotton.

4. PBS, pH 7.0: To make 1 L of 10X PBS, pH 7.0, mix 75.97 g NaCl, 12.46 g Na2HPO42H2O, and 4.8 g NaH2PO42H2O. Dissolve in distilled water (dH2O). Autoclave.

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