Chadron Biochem Day 8 – Luna
I woke up at 7:30, which was a nice break from having to wake up at 7 the last 3 days of last week because we had 8 am starts on Thursday, Friday, and Saturday. That being said, it’s certainly not as nice as getting to sleep in as I did yesterday. I texted my family letting them know I was feeling better, as I spent the weekend feeling quite homesick, but I think the return to a more structured schedule will help! At 8 I went downstairs and got my computer from the TAs’ shared space before heading over to breakfast.
My breakfast today 🙂
At 8:20 I headed over to COIL for our first activity of the day, a lesson on standard curves given by Dr.Zoetewey. We learned about serial dilutions, calculated the varying volumes of Bovine Serum Albumin (BSA) stock solution to use in our lab, and learned how to create standard curves for our data using excel.
At 9:30, we transitioned to working in the lab, where we did Bradford assays up until lunchtime at noon. My group did not have a high enough concentration of BSA during our first attempt to get a wide enough range the first time, so although we had an r^2 value of about 0.98, our spectrophotometer readings were not very consistent. After lunch we were instructed to not do the 10-fold dilution of BSA in order to create our stock, as we had done earlier in the day, and this time our Bradford assay worked much better! Our solutions actually turned blue, and we were able to find the ideal absorption range for our instrument.
Knowing our ideal range, we moved onto creating a Bradford assay with our protein, but even our highest concentration (a 1x dilution, we also had a 10x dilution and a 25x one) had an absorption value lower than our range. The other group with our same protein got similar values, so our professor said we should just run our gel for now and see if our protein concentration is high enough.
After finishing up our Bradford assays, we set up our samples for gel electrophoresis. First, our instructor put in the MW standards and demonstrated how we had to load each well using special bendy micropipette tips (aka narrow board gel-loading pipette tips). Then we loaded the four other wells assigned to our group with our induced cell sample, lysate, purified protein, and flow-through. After running our gels, for 30 minutes, we started the staining process!
One of my group members (Group 12, best for last :)) loading our final well.
We then embarked on the 10-minute walk to the dorms to change before dinner, as we have to dress nicely.
For dinner today we had green beans, mashed potatoes, chicken-fried steak (don’t ask me what this is, I could not tell you…), and some sort of white-ish mystery sauce! I really enjoyed the conversation during dinner, my table spent most of the time laughing about what names and personalities we would take on if we had to run away after committing a crime. After dinner, we met in coil for dorm meetings and a fun obstacle course to model column chromatography, gel electrophoresis, and affinity chromatography! I was very grateful to have a fun opportunity to practice what we had learned and gain a more intuitive understanding, so thank you to our professors for being such good teachers!
Finally, we ended the day with a bit of study time and a birthday celebration for Nima!
Luna is a student from Portland, Oregon who loves learning, spending time with her four siblings, and spending time outside. She hopes to pursue some combination of chemistry, biology, computer science, and engineering in college. Some fun facts about her are that she has a 1,612 day Duolingo streak and over 40 plants in her room!