Building a gene regulatory network, I think, is sort of like solving a jigsaw puzzle. Little by little, discovering which pieces go where–through logic, through the process of elimination, through experimentation. My weeks in the McClay lab have usually looked a little something like that, each day a slow and steady step towards creating something that is bigger, more complete, and eventually can tell a story.
Analyzing the relationship between genes means having to carefully select and examine the behavior of each gene. The process by which we do that takes a week, or two at most, and so my time in the lab has been a pretty steady weekly routine of pipetting, incubating, and washing. Rather than a day in the lab, then, I’ll be briefly describing a week–four days that lead up to the final day, the final reveal.
The first step that I take is creating the probe of the gene we’re looking at, which will allow for us to locate the expression of the gene. This process normally takes two days, as it requires a good amount of time for incubation. The first day involves a lot of pipetting, in order to prep the plasmid for use, while the second is the actual synthesis of the plasmid. Both are rather short days–they take only about two hours or so, as well as some waiting. While I wait for the plasmid to incubate, I usually spend the time going to lunch and enjoying the sun.
In-situ hybridizations are next, and each lasts three days. After we have prepared the well plates with embryos of different stages, the process involves a lot of washing as we prep the embryos for antibody staining in order to witness the expression change over time. These are a very systematic two days, with short intervals of waiting time and repeated activity in order to ensure all undesirable qualities are washed away. These days are longer and always busy, and I still haven’t figured out the best time to eat lunch, but it’s interesting because there’s always something that needs to be done. The last day of in-situs are the most important, though, because that’s the day of staining–the day we’re able to obtain the gene expression data we’ve been prepping since the beginning of the week. The hardest task of the day is learning at what point the embryo is done staining, and when expression of the gene has reached its utmost point. My mentor still helps me with this, but once I think it’s done, I stop the reaction. Sometimes, the wait can last hours, but I check on the embryos every 15 minutes or so. However, like the previous two days, it’s a tricky and busy day.
Sometimes, days end short or go longer than expected. But I’m glad to have this kind of routine, an understanding of what to expect, and a kind of independence that allows me to do things at my own pace, on my own schedule, and to have the authority to dictate when I can go on my lunch breaks or not. I feel a lot more respect in that sense than I did in other working environments, and have really been enjoying the past few weeks working at the McClay Lab.
Honorable mention: last Monday was a special day, because instead of the usual, my lab and I went out to the marine lab in Beaufort to collect sea urchins! It was hot and the sun was beating down strong, but we had a great time out on the sea, feeling the breeze and taking in the views. I came away with a little bit of a sunburn, and the memories of an exciting adventure.