Qualitative Determination of an Unknown Genotype of Drosophila melanogaster Through the Creation of Various Fly Cross
Prepared for
T.A. Lillian Doerfler
PCB 3063L-013
By
Hana Soueidan
Partners:
Kristina Hernandez
Brendan Meyer
Andrew Vasquez
Feb. 24, 2017
University of South Florida
Materials and Methods
Culture Vial Preparation and Drosophila melanogaster Maintenance Vials for maintaining D. melanogaster were prepared by packing dry fly food, mixed with equal parts of deionized water until the food was the consistency of mashed potatoes, into an inch-thick layer at the bottom of the vial. A 4-5 granules of yeast were sprinkled on top of the fly food. A plastic netting was folded and inserted, hovering over the food, and a foam plug closed the vial. The vial was labelled with the date, …show more content…
FlyNap, a commercial fly anesthetic, was administered to anesthetize the flies so that the vestigial adults could be obtained. Placing the vial on its side, a metal wand with a brush on its end was dipped in FlyNap and slipped between the vial wall and the foam stopper to prevent flies from escaping. The wand was ensured not to touch the walls or media. The flies were exposed to the anesthetic for 45 seconds to a minute, followed by 10 second intervals if some flies were still conscious, keeping the vial on its side. The vial was lightly tapped to knock the adult flies from the netting and onto the vial walls. Once all the flies were anesthetized, the foam plug was removed and a paintbrush was used to obtain the adults and place them on a small white board for observation under a light microscope. No live adults were left in the vial. The adult flies that were obtained were observed beneath the microscope, scored phenotypically, and separated by sex for the purposes of learning to sex the flies. These adult flies were then euthanized in the Fly Morgue. The vial was then placed back into the 25C
Prepared for
T.A. Lillian Doerfler
PCB 3063L-013
By
Hana Soueidan
Partners:
Kristina Hernandez
Brendan Meyer
Andrew Vasquez
Feb. 24, 2017
University of South Florida
Materials and Methods
Culture Vial Preparation and Drosophila melanogaster Maintenance Vials for maintaining D. melanogaster were prepared by packing dry fly food, mixed with equal parts of deionized water until the food was the consistency of mashed potatoes, into an inch-thick layer at the bottom of the vial. A 4-5 granules of yeast were sprinkled on top of the fly food. A plastic netting was folded and inserted, hovering over the food, and a foam plug closed the vial. The vial was labelled with the date, …show more content…
FlyNap, a commercial fly anesthetic, was administered to anesthetize the flies so that the vestigial adults could be obtained. Placing the vial on its side, a metal wand with a brush on its end was dipped in FlyNap and slipped between the vial wall and the foam stopper to prevent flies from escaping. The wand was ensured not to touch the walls or media. The flies were exposed to the anesthetic for 45 seconds to a minute, followed by 10 second intervals if some flies were still conscious, keeping the vial on its side. The vial was lightly tapped to knock the adult flies from the netting and onto the vial walls. Once all the flies were anesthetized, the foam plug was removed and a paintbrush was used to obtain the adults and place them on a small white board for observation under a light microscope. No live adults were left in the vial. The adult flies that were obtained were observed beneath the microscope, scored phenotypically, and separated by sex for the purposes of learning to sex the flies. These adult flies were then euthanized in the Fly Morgue. The vial was then placed back into the 25C