On the first day of phage hunting, upon entering the lab for the first time, I was expecting to see the materials already set out for me, and a lab notebook with instructions neatly lying in the middle of my personal lab bench. Instead, all I see is a Bunsen burner which having always scared me to death, did nothing to alleviate my anxiety.
If I were to describe phage hunting in two words back then, it would have been Unexpected Freedom. On that first day, we were given a quick outline of what we were supposed to do, and then proceeded to be let loose in the lab like a pack of clueless college freshman.
The shock only lasted a few more classes, though, thankfully, and by the fifth day or so I had learned how to navigate around the lab, carry out the various protocols with ease, never flip my plates to prevent the agar from shifting, and bond with the Bunsen burner to make the most out of my aseptic technique. All my blunders and contamination problems in the beginning of the semester diminished and it was smooth riding from there. I had found my flow, and felt empowered and unstoppable in my white lab coat.
The following days can be summed up by the following:
Enrichment? Check…1st streak? Check…2nd streak? Check…3rd streak? Check…4th streak, Check…MTL? Check…Web Plate? Che- Aww crap…
And that’s when I hit the first of my biggest walls. The 6 web plates that I made to flood and collect as my HTL (Highly concentrated stock of phages) came out poor in that there should have been a gazillion plaques on each plate, but they only had about 15. In the end, it took me two more tries to finally obtain my HTL.
But the pride in my HTL evaporated when I realized that there was more work to be done in isolating its DNA. It took me five tries of tedious washes with isopropanol and centrifuging to obtain a high enough concentration, and that was the class right before final presentations, another stressful time.
While I thought my problems ended after giving my final presentation, immediately after I learned that my DNA was not refrigerated. So, as of now, my struggle in Phage Hunting continues as I prepare myself for another round of DNA prep and, considering that I have ran out of HTL, need to work desperately to make this last one count.
So what are my thoughts on Phage Hunting three months into it? Despite all the work and challenges, Absolute Delight. Working in the lab and being empowered with a Bunsen burner and a pipette relieves the stress from school and studying all the time. But most of all, after reflecting on my achievements, seeing how far I’ve come, and finally seeing my phage via electron microscopy, I am certain that I will never regret taking Phage Hunting.