Drosophila Egg-Laying Preference Assay: A Method to Test Decision-Making in Flies
DEŞİFRE METNİ
– To test a female fly’s preference for egg-laying, or oviposition sites, provide an individual female fly with a choice between two different substrates on which için deposit her eggs within a test chamber. The fly will compare the available options and choose the sites she prefers, usually the substrate best suited for her offspring.
Ensure the fly is primed için lay many eggs, so there are robust egg numbers videodan each trial. At the end of the experiment, remove the fly, and count the eggs on each substrate için determine the fly’s oviposition preference.
In the example protocol, we will use a high-throughput assay için test egg-laying preference for plain or sucrose-containing agarose substrates.
– Devam et with the experiment by obtaining previously constructed acrylic egg-laying chambers. Insert plastic sheets into the top, loading a piece of the chamber için serve as the bottom surface.
Next, anesthetize the females on a carbon dioxide pad, and load them individually into each egg-laying arena. Allow the flies 30 minutes için recover from the carbon dioxide and için acclimate için the new environment. While the flies are adjusting, fill a tube with agarose up için the 10 milliliter mark. Then, prepare a 150 millimolar sucrose substrate by placing 750 microliters of 2 molar sucrose solution into the tube. Keep a bottle of melted 1% agarose videodan a water bath.
Prepare the plain substrate videodan the same manner, but replace the sucrose solution with distilled water. Next, obtain the substrate piece of the chamber and pipette 1,000 microliters of the agarose substrate into each trough. Set the agarose aside için solidify for 30 minutes.
Once the agarose substrates and flies are ready, assemble all three pieces of the egg-laying chamber and then take out the plastic sheets. Place the chambers videodan fly incubators overnight.
Anesthetize females by injecting carbon dioxide into the chamber. Disassemble the chamber and discard the anesthetized flies. Finally, take pictures of the eggs for record-keeping.
