by Eleni Padden
So here I sit in my dimly lit dorm room, listening to the ever-present rain patter down outside, just enough to annoy the bejesus out of everyone on campus but not enough to actually cause class cancellations. Alas. The weather is strikingly reflective of my current mood, having just returned from the lab. It’s been a long few weeks in there, plating, enriching, diluting, measuring infinitesimally teensy amounts of potential phages into centrifuge tubes, etc. I must say, things could be going better. Here’s a quick-n-dirty synopsis of what’s gone on with my valiant but ultimately fruitless attempts to isolate a phage (elusive little buggers) in the past several weeks:
-Week one consisted predominantly of collecting samples from around campus. A few friends and I ventured down to a particularly janky looking stream about ten minutes away from the freshman quad (we’d heard through the grapevine that M. smegmatis has a thing for moist materials). We gathered our samples in some 15 mL test tubes, labeled those pups, and kept em cool in our fridges. My roommate was briefly concerned at finding dirt filled tubes in the fridge, but I told her it was in the name of Science, and all was well. The next lab time we had, we spent practicing honing our aseptic technique skills and slowly/awkwardly executing our first direct-plating of samples. We also prepared an enrichment culture in a flask, which we’d deal with at a later date. We had to let it ‘stew for a while’, if you will.
-The next few labs consisted of many emotions and a lot of direct plating. I experienced what we’ll refer to here as some ‘technical difficulties’– long story short, I plated my samples upside-down. Once again, awkward. Upon seeing my own personal results of absolutely zero phage on my petri dish, I knew I needed to try out my enrichment sample. I diluted it from a 10^0 concentration all the way to a 10^-4 dilution, with 10^0 containing the least diluted sample (this was, in fact, entirely undiluted). In theory, the contents of my 10^0 microcentrifuge tube would, once transferred to a petri dish, yield the dish with the most phages since it was the least dilute of all the samples. But once again, disappointment ran rampant in the lab. Very few people got viable phages from either their enrichments or their direct platings– myself being one of the forlorn majority. However, science does not sleep. We all pressed on.
-I kept with the method of direct plating for the remainder of my phage-isolation attempts because it was simpler and I could accomplish plating many samples during one lab session… and in retrospect, probably also due to a subconscious desire to redeem myself for the initial upside-down plating. I collected various new environmental samples and essentially started at Square One. This seemed nausea-inducing and daunting at first, but as it happened, I came to realize I’d gotten a lot more comfortable with lab proceedings as the time had passed, and, as always, I pressed on. I directly plated three new samples (one which was annoyingly ornery and wouldn’t separate out simply with the phage buffer and a little vortexing; Dr. Schildbach and I had to do battle with it, ultimately resorting to the centrifuge machine). After plating the new samples and coming back the next lab session, I was pleased to see– FINALLY– some evidence of a phage. Beauty incarnate.
-Streaking: belligerently running naked on public grounds OR a scientific method of phage isolation having to do with rubbing sterile wooden sticks in differing concentrations of lines on an agar plate. Luckily in the lab, we did the latter. After seeing some possible phages on my #4 plate, I streaked the plaques out and poured a mix of toasty top agar and .5 mLs of M. smegmatis on top of the newly streaked petri dishes. I collected several new environmental samples, just in case, and direct plated those guys as back-ups in case #4 ended up letting me down. The next lab session resulted in more signs of hope, as it seemed like #4 might actually have a phage (the streaking had questionable but nonetheless possible success)– and all three of my environmental samples had minuscule punitive plaques growing. It was with great joy and rapture that I streaked from various spots on my #4, # 6, and # 7 plates, hoping for evidence of true phages upon my arrival to the next lab sesh.
-This leads us, dear reader, to where I am now. I jauntily walked into lab today, hopes sky-high, and went to go check out my dishes. Nada. Nothing. Zilch. Not a phage in sight. All of the streaks had failed me miserably (or had I failed them? No, no, don’t blame yourself Eleni… hostile is the nature of science). In all seriousness, though, none of the phages ended up working out, and now I’m back to the dreaded Square One for a second time. Today I attempted another hail-Mary streaking of my original #4 plate, just in case. I’m also the proud parent of a newly adopted baby phage, a boy, named Zachary. Kidding, phages aren’t actually alive and thus can be neither babies nor boys (although I guess I could eventually name it Zachary). My wonderful lab partner, who has had lots of success with her phages as a result of quite a lot of skill and a dash of luck, donated one to me. I streaked it from four different locations today and hopefully, if there is a God, I will have a real, not-live phage come next lab. Please keep me in your prayers/wishes/etc.
For now, this is Eleni Padden, signing off. Have a phagetastic weekend! (Yeah, there weren’t nearly enough phage-related puns in this post so I felt like I had to throw at least one in for good measure). Adieu! Ü
P.S. Check out my sweet photo-montage: