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Unformatted text preview: Drosophila PCR Laboratory Procedure Extraction of DNA from Drosophila 1. Students will work in groups of 2. Each group will get one fly ( Drosophila melanogaster ) for analysis. Each fly will be from a sample of the mixed population vial of flies (Bar eye flies and black bodied flies) or a control fly from a pure breeding stock vial (either Bar eye or black body). Flies will be anesthetized and assigned by the instructor. 2. Label one 1.5ml microtube for each fly with a unique mark (initials or numbers). Place one anesthetized fly in each labeled tube (one per group). 3. Add 50 l of homogenization buffer to each tube and grind the fly with a plastic pestle. Grind the fly as completely as possible. Use care when grinding and when removing the pestle to leave as much fly homogenate in the tube as possible. 4. Incubate the tube for 25 minutes at 30 C. 5. Incubate the tube for 1-2 minutes at 95 C or in a boiling water bath. 6. Briefly spin (pulse) the homogenized flies in a microcentrifuge so that remaining tissues form a pellet at the bottom of the tube. Balance your tube with that of another group. The fly DNA should be in the supernatant. Place this sample on ice until the PCR reaction is ready for your DNA....
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