By Kathleen Kranzlin
Almost as soon as Phage Hunting began, it seemed that the recurring theme would be repetition. I had to repeat two steps in particular: streaking, and performing the Titer Assay. Therefore, I can confidently say that I am now an expert on these two techniques. The multitude of repetition also served another purpose: it ultimately helped me decide my phage’s name: Persnickety.
The first couple of weeks of Phage Hunting went well. I found my phage among the dirt procured at the community garden and I proceeded to the next step, streaking. Being the naive and unfamiliar lab student that I was — and still am to some degree — I had no idea what I was getting into. After almost an entire month of streaking, which included extra lab time multiple times a week to make up for being behind, I finally managed to isolate a single plaque morphology. Interestingly enough, none of my streaks were riddled with bacterial contamination. Instead, my phage refused to behave; they always merged together and resisted isolation. How many streaks did I do in total, one might ask. I had streaked eight times, with each set of streaks including streaks of at least two plaques. Nevertheless, I had finally procured my three solid streaks: T-streak #3, T-streak #5, and T-streak #8.
Finally, I could move on to the next step, but luck was not on my side. After suspecting that my first Titer Assay would result in contamination, I was advised to perform a second one. Two days later, I took out my plates from Titer Assay 1 and Titer Assay 2 and proceeded to laugh. Once again, I would be repeating the Titer Assay. The first TA resulted in contamination, much like what was suspected. The second Titer Assay had really weird results; the only plates with plaques were my 10-3 plate (which had two) and the 10-5 plate which had one and all the other plates had a lawn of bacteria. So, like streaking, I went back and repeated the Titer Assay. Two days later, I was once again laughing at my luck. The plates from Titer Assay 3 were half cleared, and showed no sign of clear plaques. Once again, I was back to performing the procedure that I had become all too familiar with. However, in the next lab period luck was on my side. Not only did I have a beautiful Titer Assay, but I had also found a perfect web plate, 10-2. Thankfully, after a little over two weeks, I could move on to finding my Medium Titer Lysate.
For the rest of the procedures until Phage Olympics, my phage behaved and I did not have to repeat any of the steps with the except of DNA purification—my first DNA only had 86.3 ng/μl. When it came time for me to choose my name, I looked back on my lab time and realized how fussy my phage had been in the beginning steps. Therefore, the name Persnickety was born. With luck now on my side, I was able to finish on time (thanks to some extra lab sessions) and Persnickety competed in the Phage Olympics.