Materials and Methods

Bottles were used to prepare crosses between male and female Drosophila. Instant Drosophila Medium Formula 4-24 and distilled water were added to the clean bottles. Fleischmann’s Active Dry Yeast were sprinkled on top of the medium. Virgin male wild-type Drosophila were carefully added to two of the bottles. Then, wild-type females were added. The last two bottles were filled with male and female white-eyed Drosophila. The bottles were set aside for ten days to ensure that the mating cycles completed a supplemental amount of virgin Drosophila in various life stages for collection.
In order to collect virgins, the adult Drosophila were disposed of in the morning via dumping them into Fly Morgue. Those that remained were pushed into the medium so they would not affect the outcome of the virgins later. Smaller amounts of food and distilled water, along with yeast, were put in small, clean vials. These vials were labeled male w+, male w-, female w+, and female w-. After 8 hours, the adult virgins that hatched were put to sleep using FlyNap, collected and separated between both sexes of all types using a microscope. Curly flies were removed and disposed of in the Fly Morgue, then males and females were separated and placed in their appropriate vials. Collections were made three days each week for two weeks.

Three weeks later, mating assays were observed by crossing male w+ with female w+, male w+ with female w-, male w- with female w+, and male w- with female w- and recording the time it took for them to copulate. Time was recorded in intervals of 20 minutes, and the Drosophila were disposed of in the Fly Morgue afterward. Multiple assays were performed throughout a time period of two weeks when adequate amounts of Drosophila for each type were present.

**This entire procedure was conducted a total of two times, having four weeks apart, in order to accumulate a reasonable amount of Drosophila for the experimental data.