Emily Guo, Week 3, All About Cell Culture

This week, my grad student has been teaching me cell culture protocols, how to care for suspension cells (Jurkat) and adherent cells as well as how to prepare cells for DNA isolation. This will be very helpful for her as she spends a few hours everyday maintaining the cells and making sure they do not become overcrowded, which would lead to wonky expression. As we are also analyzing cell expression, this would be a problem. We’ve also been doing serial dilutions, which are important for isolating individual cells and purifying samples. The samples from before I came to the lab - which were cut by directly introducing Cas9 and gRNA - had a few candidates whose gels had indicated that the samples loaded needed to be purified. By doing some careful calculations and diluting the cells, we can deliver 1 cell to each well and grow wells of identical cells, effectively separating them out.

Cell Culture Room

My grad student has been spending a lot of her time preparing for a talk about the research she was conducting in the lab, which she gave on Tuesday. It was a lot of work and very stressful, so the beginning of the week has been a bit slow. However, everything turned out well and we went to Antika to get pizza and to celebrate. 

The next step will be to create plasmids for another gene (for which the inserts are already created), which we will be starting next week. Anna and I will both work on the project and hopefully grow successful E. coli after the first plating, now that we know what conditions should work. We also finally received the last set of primers for our qPCR, which will be conducted on Monday. qPCR with cDNA synthesis allows us to analyse the expression of our cell lines (via mRNA) by converting the mRNA into cDNA, then allowing us to see the PCR amplification of the DNA in real time. 

Isabella, another lab technician, also returned from her two week vacation. Her first day back, I saw her and thought I had come to the wrong lab… Our lab has also grown to include two more students, Lucas and Noah, who are working with Cathy and Martha; Lucas is also a rising senior in high school. The size of the lab has almost doubled with the addition of us students! Arun, who used to work with Alyssa on her project, dropped by to have a chat. I tried to sneak in a panorama of the lab during lunch when no one was looking:

The lab now takes up two tables at Strokos.

Meanwhile, I’ve found a new supermarket on 160^th that is both cheaper and has a wider selection of goods. Unfortunately, the trip back entails scaling an increasingly steeper incline, which is made worse by a backpack full of groceries. But I will prevail!

I’ve also bumped into a Peddie student on my way to the lab one morning. I'd recognized her face in the crowd commuting to work two times before, but the first time I'd thought that I had just misidentified a familiar-looking stranger. The second time, however, she was wearing a Peddie shirt, so I was determined to catch her the next time I saw her. It was nice to chat with another person from Peddie, even if we only recognize each other by face rather than by name.