My time in the Volkan Lab consists of primarily two procedures, PCR/running gels and ChIP (chromatin immunoprecipitation). This means that my day is usually split into half collecting/dissecting fruit flies and pipetting for these two procedures. I usually start my day by collecting Ir84a mutant male flies (we only study gene regulation in males so they have to be separated from the females) and then preparing buffers so that I can start ChIP by moving onto dissecting previously collected males. Depending on how many flies were previously collected, I will either do an antenna or a head dissection which take roughly 90 and 30 minutes respectively. Once the dissections are done I proceed on with the next steps of ChIP over a course of a couple days. These steps include fixing, washing, homogenizing/sonicating, immunoprecipitation and qPCR. This process of ChIP is the primary tool being used to test the hypothesis of my project: Ir84a mutants will have the chromatin wound more around the fruitless gene than wild-type flies.
In addition to ChIP, I also make and run a lot of gels. The primary purpose for which I run the gels is to confirm that the lines of flies I am using are indeed mutants for the receptors in question. For example, I am studying flies that are mutants for the Ir84a receptor and so I perform PCR on these DNA samples with primers for the Ir84a gene. If a band is seen on the gel for this Ir84a primer, then the line was not actually a mutant line, but if no band is seen, then the line is indeed a mutant.