Ever since I saw this self-care YouTube video by some random woman saying it’s ~self-care~ to wake up early enough so you don’t have to rush to get ready + have some time to yourself, I’ve woken up at 7-7:30AM everyday- minus weekends. So, I start my day off at 7 and go into lab around 10-11AM depending on the day. We have several back-to-back imaging days, so I tend to stay by the BM3-BC Colony Processing Robot, or as I call it, the phenotyping robot, as it takes images of plates with growing crypto. in it. While the robot does its thing, I take note of the start and end times and identify deletion strains from previous sets of plates that have already been imaged. I identify these strains using Fungidb which is a nice deletion library that has an extensive description of each strain and contributing papers. Occasionally I’ll run into several hypothetical proteins which is exciting because it gives an opportunity for an additional project to dig deeper from what we’ve found to learn more about the strain. After imaging of a set of plates is complete, which can take 50 minutes to 2 hours depending on the size of the set, I mark for growth on plates with 32 or greater micrograms per milliliter of FCN. I don’t find too many, as 32 is already a generally high concentration. Sometimes I think about the gap between 32 μg/ml and 64 μg/ml and wonder if it’d be worth repeating this experiment, only zooming into those concentrations. We’ve only seen two strains that have been able to grow at 64 μg/ml, which although that’s a good thing, it’s still kind of threatening and scary to me as 64 μg/ml is such a large concentration. I’d definitely like to study more about those two strains as well and its genes to learn more about how it’s able to withhold such high concentrations of FCN. The data inserting process will take much of my time this week because I’m observing so many deletion strains (~4700) and I’m also learning how to use Excel, so that’s fun. Later this week I expect we will begin the process for confirming the FCN resistant strains.
Here’s a visual to understand what I mean a bit better:
This is a plate with a concentration of 64 μg/ml, and there is growth in position F1. I’ve identified this to be a deletion named arginine biosynthesis by the Fungidb deletion library.