Take a Walk in My Lab Shoes

What’s up readers? This week I’ll be sharing a typical day’s routine from my time in the lab. My days start at 9 in the morning when I usually get to lab. I like to bike to work in the mornings from my apartment to wake myself up. My morning schedule completely depends on what experiments I ran the night before, but usually my mornings consist of completing Western Blots or harvesting RNA from my cells. Western Blots require an overnight antibody incubation step, so on days where I run Westerns, the next morning’s first step is to complete it with the necessary washing, secondary antibody incubation, and image development. On the mornings that I’m not developing blots, I often have RNA harvesting to do. One of my most common experiments is comparing RNA levels between different cell treatments using Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR). These experiments often require treating cells with certain drugs, some that inhibit our kinase of interest, others that inhibit other kinases, and others that act as control drugs. These incubations are often 24 hours, but can be as long as 72 hours depending on the drugs used. On days that I harvest RNA, I usually set up the RT-qPCR experiment. This involves an intermediate step known as the cDNA synthesis which converts the sample RNA into DNA for the PCR reaction using reverse transcriptase. After this I set up the RT-qPCR reaction. The entire procedure takes about half the day to set up, but it yields really good data. By the time I’ve finished developing my blots or setting up an RT-qPCR, it’s usually around lunch time. Most of the time I pack a lunch, but sometimes I’ll head to the Hospital Cafe or an off campus restaurant for lunch with some other members of the lab. If I eat in the lab, I like to analyze data or read papers during my lunch. I’ve found that there’s always more to read, and especially since my project is relatively open-ended, there are lots of papers out there that would aid my search for new mechanisms of Abl kinase mediated metastasis. After lunch is usually when I finish my tissue culture work for the day. I keep around 6 cell lines growing at all times, and to keep these cells healthy and happy, I have to dilute them for growing space and replace the media which they grow on every other day. Most of the lines I keep are lung cancer lines, but I also keep a line of 293T cells for the virus experiments that I occasionally do. In addition to regular upkeep of these cells, on most days I need to set up cell experiments. Often these are drug treatments, but they also include virus transductions, cell viability screens, and trans-well migration assays. For these other experiments, I have to count the cells that I have using a (very carcinogenic) dye, calculate the correct dilution ratios, plate a precise number of cells, and if I’m applying drugs, dilute the drugs to appropriate amounts. My tissue culture work can take as little as 30 minutes when I only need to split my cells, but it can take upwards of two hours on particularly busy days (usually fridays). After tissue culture, it’s often time to call the day quits. But if there’s extra time, I will usually run a Western Blot gel of do a quick cDNA synthesis to get a little ahead for the next day. I’ll usually bike home from work, relax for a little, cook dinner, and head to bed to rest up for the next day! My days can be pretty diverse depending on the experiments I need to complete. I love the variety of my schedule. Each day brings new challenges and new data to show to my lab-mates and to put on my poster. Although the 9 to 5 research life is long, I love my days spent in the lab. I’m nearing the end of my summer research, so in the coming weeks I’ll be finishing my research and preparing my poster. Can’t wait to check back in next week!

-Brennan

Leave a Reply

Your email address will not be published. Required fields are marked *