A normal day in the lab usually begins with checking on my 293T cell lines to see if they are confluent enough to passage to a new plate. If the cells are 80-90% confluent, then I will thaw my media and trypsin, prepare the cell culture hood and then passage 10% of my cells to a new plate so that they don’t become overconfluent and die. I usually need to split my cell lines every other day.
After I do this, sometimes I will have a mini prep or a midi prep of bacteria with my designed gRNA plasmids culturing in the incubator overnight, so I will take that out of the 37° shaker. I will begin the protocol by centrifuging the tubes with my transformed bacteria for 10 minutes, so during this time I check in with my mentor. We will go over the procedures I will need to do that day and why they are necessary for my project. This is usually a good time for me to ask questions about the various protocols for the day’s tasks and be sure I understand each step. After this check-in, I carry on with the mini prep, or whatever procedure I need to complete first for the day.
By the time I am done with the day’s first assignment, it is usually time for lunch. Some days, the lab will all go out to lunch or someone will order pizza for everyone. These are the best days because I get to hear about the cutting-edge research they do and also get to know them outside of the lab setting. After lunch, I will complete the rest of my tasks, whether that includes transfecting cells, changing media, harvesting cells, PCR, restriction digests, surveyors or designing new guides for my project.
Before I leave for the day, I will transform one of my plasmids into competent cells or start a mini prep from bacterial plates that I have already prepared. This is done at the end of the day so that the bacteria can grow overnight. Overall, the lab is a relaxed, collaborative environment where I don’t feel afraid to ask questions.