Each day in lab can easily become very hectic and I often find myself learning new procedures, having interesting conversations on topics I’ve never heard of, and meeting new people every single day. However, a pattern has begun to emerge in my weekly schedule as I have begun to perform similar experiments multiple times in the past few weeks. For instance, I use a particular line of ‘immortal’ human cells called 293T cells daily for my experiments. To prep these cells for experiments, I must go through a 3 day process of plating these cells to grow, transfecting them with a desired plasmid, infecting them with a virus, and harvesting them to be tested in various ways. I usually perform this cycle two times a week and find my harvesting days, often the days that I also perform experimental tests, to be the most exciting. My most recent harvest day, Friday, was yet again a fairly exciting day:
On Friday, I arrived at lab a bit after 9 and placed my things at my desk, talking to the two postdocs with desks surrounding mine as I settled into lab for the day. Then I went to the tissue culture room in our lab to harvest my cells that I had been preparing over the past two days. This procedure takes about an hour in total, involving checking the cells to confirm they’re living, removing the cells from the plate, dividing the cells into smaller test tubes for my two different experiments for them, a luciferase assay and a western blot, pelleting these cells so they can be stored, and plating new cells to be used in my next round of experiments.
After this, I met with the principal investigator in my lab, Stacy Horner, to talk about my project and help prepare for my chalk talk in the following week. She was able to guide me on the next steps in my project, as well as give me advice on teaching on the lab’s research and presenting in general for my chalk talk. After this meeting, I went back to my lab bench to conduct experiments on the cells I had harvested previously. I began with the luciferase assay, a procedure that measures interferon induction in the cell through tagging with a fluorescent substance, luciferase. This procedure usually takes around 1-1 ½ hours to set up. After this assay was finished, I constructed graphs on my computer to display the data and average it with other replicates of the same experiment. Then I used the rest of my harvested cells to perform a western blot, which helps to visualize whether or not a particular protein is being expressed in the harvested cells. This procedure takes a few hours, requiring quantification of the concetration of protein, separation of protein by size on a gel, and identification of the protein with antibodies. All throughout these experiments, I was talking to my labmates about various subjects in lab as well as other fun and less lab related topics. When this experiment was completed, I ended my day and headed out of lab around 5.
