Busy Days
Table of Contents
Sunday, March 9
163.8 lbs (Day 72, starting from 162.6 lbs on Dec 28) | 🏃🏻 Done (6/6) | ★★★★
The weather is absolutely amazing. I had a morning gym session and worked on slides for a presentation while finishing some remaining experiments. I try not to go to the lab on Sundays, and although I had never done this in my Master’s in Korea, now that the campus is so close, I sometimes feel like going if I don’t have any appointments. Even though I went to the lab on Sunday, I didn’t mind too much because the weather was insanely great.
Following Friday’s experiment, I performed one more reverse Ni purification for the E-to-C mutant protein and discovered that the issue was in the reverse Ni step. The band intensity of the sample right after TEV cleavage was good, but it dropped significantly after the column. This mutant might be unstable on the Ni-NTA resin without his-tags, so I decided to focus on the labeling experiment for the M-to-C mutant. After dinner at home, I headed to the Hub, read my journal club paper, and made some outlines.
Monday, March 10
165.0 lbs (Day 73, starting from 162.6 lbs on Dec 28) | 🏃🏻 Rest (0/6) | ★★★★
Outside of Work
Yesterday, I had so much homework that I ended up sleeping around 2 am, so I decided to skip my morning workout. I can’t imagine how people who exercise every day manage it while doing their own job too. The final week of Reaction Mech has begun. After three classes on Cross Coupling, there’s a final exam next Monday. And I just knew that, surprisingly, it’s an open-book exam😶 I think this is the first time I’ve ever had an open-book exam in a class like this. It only happened once(10 years ago…), when I had a final for a college general education course where I had to write an essay within a set time. Either way, I’m glad I don’t have to worry too much about studying. What’s even more amazing is that TAs even give us a chance to correct our final exam! Of course, it would be better if you aced it on the first try, but still great. Today, I got off work around 7, had dinner, relaxed a bit, and got a good sleep.
Lab Work
I’ve now finished the protein prep, and I’m starting the experiment to see if the compound I synthesized reacts with the protein I isolated (i.e., whether they form a covalent bond). But the major problem was that the LC-MS needed for the experiment broke down again😵(why does it happen one week before my exitalk), so after dealing with a lot of hassle, I ended up using one from another lab (I’ve reserved it for tomorrow). For labeling experiment, there are so many factors to consider. Time, concentration, buffer composition, temperature and more that I consulted with some experienced people from another lab to set the conditions. I really hope the results turn out well.
Tuesday, March 11
165.4 lbs (Day 74, starting from 162.6 lbs on Dec 28) | 🏃🏻 Done (1/6) | ★★★
Outside of Work
Today, after a great workout, I focused on my experiments. To be honest, I skipped the QBC journal club again. I have one more chance to skip it, so I guess I’ll skip it again next week. There is just too much to do, and unless someone from my CCB cohort is presenting, I don’t really feel the need to attend. The experiment wasn’t going well, so I went out like a light as soon as I got home.
Lab Work
For the labeling experiment, I used an automated multichannel pipette for the first time ever, and it was absolutely amazing. To be honest it wasn’t necessary because I only had 16 compounds that I made, and with the positive and negative controls, there were just 18 samples in total. But still it was useful. Needless to say, for screenings where the number of samples increases exponentially, this tool can be essential. It’s always so satisfying to learn new equipment and techniques.
After letting the labeling reaction run at room temperature for two hours, I spent the remaining time preparing my exit talk presentation slides. Since my English isn’t great, I’m trying to make the presentation as engaging as possible by using detailed figures and animations, which is really time-consuming. I wish I had started it sooner🏃🏻
Later that evening, I went to use the LC-MS at another lab, but then I noticed that my protein doesn’t seem to be visible! I thought maybe I had done something wrong, so I checked a sample from a friend who booked the LC-MS with me, and it showed a weird pattern too. We spent two hours troubleshooting and there were moments when I felt as thrilled as in that video when everything suddenly got fixed, haha, but in the end, we still failed😞 So, I stored the sample at –20°C and decided to look for another LC-MS tmr.
Wednesday, March 12
164.4 lbs (Day 75, starting from 162.6 lbs on Dec 28) | 🏃🏻 Done (2/6) | ★★★★
Outside of Work
I had a refreshing workout, then grabbed a coffee with a grad student in the lab. He explained in detail how to make figures, and I’m really thankful for that. I was especially surprised to learn that we can adjust gel images on a laptop using software called Image Lab instead of manipulating the image in the machine. It was shocking that you can also do quantification based on band intensity!
After that, I attended class, and there were no special events. When I got home, I did some meal prep. I worry that I might miss out on the most important experiment if I can’t find a solution for equipment issues.
Lab Work
I managed to use an LC-MS in another lab. It looked fine when I only checked proteins! I spent the whole day talking with my mentor while focusing on making my exit talk presentation. In the afternoon, I attended my mentor’s group meeting presentation where he also shared the background of the rotation project I am in charge of. It was interesting to listen to. It seems most postdocs handle two or more projects. If both are very demanding, it must be really hectic. On top of that, people with children also have to take care of their families. There are just too many things to do at once in this world🫠
Thursday, March 13
164.9 lbs (Day 76, starting from 162.6 lbs on Dec 28) | 🏃🏻 Missed (2/6) | ★★★
Outside of Work
Yesterday, I went to bed a little late and ended up skipping my workout. It always seems like there isn’t enough time when something important is coming up. I know that if I went to sleep on time, I would be more focused and work more efficiently. But the worry of not having enough time makes me keep working until I feel I’ve done enough, even if that means going to bed late. Today, without any classes, I spent the day doing experiments and preparing for my presentation.
Lab Work
I thawed a sample that I had frozen on Tuesday and checked LC-MS, but my protein wasn’t showing up well. I wondered what the problem could be. I thought that maybe freezing and then thawing it might cause the issue, so I repeated the exact same experiment. However, the protein still didn’t show up well. Then I considered that there might be a problem with the protein itself. I checked the LC-MS of the protein(just with buffer), and it looked a bit strange compared to the first time. So, I suspect that my protein might be unstable at room temperature. I plan to thaw a new aliquot tomorrow and try to check it again.
Friday, March 14
165.0 lbs (Day 77, starting from 162.6 lbs on Dec 28) | 🏃🏻 Done (3/6) | ★★★☆
Outside of Work
I managed to complete my workout and attended the “Research in Progress” class in the morning, followed by my final “Reaction Mechanism” class in the afternoon. The rest of the day, I worked on my experiments. One of my colleagues had a birthday, so around 7 PM I left work and had a light beer at our dorm. Most of my friends went out dancing at a club afterward, but I knew that clubs weren’t really for me. Also, I was feeling overwhelmed by the pressure of next week’s Exit Talk, so I stayed home to prepare and went to bed early. Tomorrow, I have to go to the lab again.
Lab Work
When I checked the newly thawed aliquot, the protein signal looked fine(although the machine had some issues with MaxEnt, which kept crashing. Actually, the next day, I discovered that the LC-MS was also having problems). So, I concluded that the protein might be unstable at room temperature. Therefore, I did an overnight labeling at 0°C, and since I didn’t have much time left for experiments, I decided to do Differential Scanning Fluorimetry (DSF). It was exciting to learn a new experiment, but I felt a lot of pressure because today was officially my last day. The results for the wild type turned out well, but the mutant came out strangely. Since I have to check the labeling results tomorrow anyway, I plan to increase the concentration of the mutant protein and try it again.
Saturday, March 15
164.0 lbs (Day 78, starting from 162.6 lbs on Dec 28) | 🏃🏻 Done (4/6) | ★★★★☆
In the morning, I went for a workout with a friend. Lately, this friend is doing really well and might even graduate early. It’s a bit sad because that means we’ll only have one more year together. Still, watching a friend succeed according to plan is one of the most rewarding feelings, so it makes me happy too.
Right after our workout, we went straight to Costco. My friend said that one of the tires was completely flat, so we filled it with air at the gas station. At Costco, I bought some new things like poke and tteokguk, and I also picked up chocolates to hand out at the Exit Talk (honestly, no one I know dislikes Ferrero Rocher haha).
After coming home, shower, and having a great meal, I returned to the lab to check the labeling results. They looked strange. I reviewed the data step by step and realized that the LC-MS (specifically the protein column) might have gone bad. I hoped that our lab’s LC-MS would be fixed quickly so I could get the data before the Exit Talk.
The DSF results using a higher protein concentration were good. My protein did not seem unstable at room temperature, and its cap-binding affinity was observed. One thing is clear: once our lab’s protein LC-MS is back to normal, I will need to run the test again. After getting home, I worked hard to prepare for my presentation.
