Like I have mentioned in my previous blog posts, my experiment is based around seeing what genes effect sexual behavior in the brains of flies. Depending on what gene (fruitless, doublesex 1 or 2, or ChAT) and what epigenetic marker (H3K27ac) or a proxy of transcription (RNA Polymerase II) we are focusing on, my day in the lab may be different.
For example, if we are just starting to look at one of the genes and epigenetic markers listed above, I will either collect adult male flies in groups of 30 or isolate fly pupa into their own bottles (1 pupa to one bottle). These collections set up group housed or isolated flies for my experiment.
Those flies are kept in their environments for about 5 days, so across that span I can collect more flies or start dissecting flies from previous collections. For dissections, I take the head off of the body of the fly and remove the mouth.
After dissections, ChIP protocol is followed to isolate the DNA that is related to the epigenetic marker we’re looking at. Following ChIP, we use computer analysis to quantify the enrichment of the respective protein or mark that the flies received from their respective environments at the promoters for each respective gene of interest.
This cycle is essentially repeated over and over again, so whatever stage of the cycle I am at is what I do during a day at lab. If I had to choose, collecting and dissections are my favorite parts!