General Supplies and Reagents

1. Baskets with 80 |im Nitex mesh (Sefar America, Kansas City, MO) for dechorio-nating embryos. To make a basket, cut a 1-in. length of a 15- or 50-ml Falcon tube at the cap end. Use the cap to hold a small piece of mesh in place. Place the basket in a small beaker to hold solutions.

2. Fine forceps, tungsten needles, and glass plates or depression slides for dissection.

3. Apple juice/agar plates: Mix 90 g of Difco agar with 3 L of water, autoclave for 50 min and cool in a 60°C water bath. Mix 1 L of apple juice with 100 g of sugar and heat to 60°C to dissolve. Combine agar/water and juice/sugar mixtures, stir, add 60 mL of a 10% solution of p-Hydroxy benzoic acid methyl ester (Sigma) in ethanol and pour the plates. The tops of 35 x 10-mm plates (Falcon) will fit fly food bottles from Applied Scientific.

4. Egg-laying chambers. To make chambers, cut a hole in the side of a dry fly food bottle and stuff it with a cotton ball. The apple juice/agar plate will fit on the mouth of the bottle.

5. Yeast paste: Mix granular yeast (Sci-Mart, Inc., St. Louis, MO) and water into a smooth paste.

6. Drosophila Ringer's, 1X (DR): 130 mMNaCl, 4.7 mM KCl, 1.9 mM CaCl2, 10 mM HEPES, pH 6.9. Make up as a 10X solution and store at 4°C.

7. Phosphate-buffered saline, 1X (PBS): 137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 2 mM KH2PO4, pH 7.4. Make up as a 10X solution and store at room temperature.

8. Fix: 4% Paraformaldehyde (Sigma-Aldrich) in PBS. Heat to dissolve. Store at 4°C for up to 1 wk.

9. Microcentrifuge tube rotator (e.g., Labquake/Thermolyne no. 400-110, available from VWR).

10. Fluorescence microscope equipped with fluorescein, rhodamine, and ultraviolet (UV) filters, bright field/differential interference-contrast (DIC), and a camera.

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