Phage or not a Phage…That is the Question!

Walking across the beach on campus I could clearly see straight ahead of me just what I was looking for.  It sat poised, ready to join me on a epic journey whose end was unknown.  Step by step I got closer to it, but it did not stagger away, it stayed as if it knew just what was going to occur.  I sat beside it, examining it closer, and then removed what I needed: a plastic knife and a test tube.  I dug my knife into its surface, carefully but with enough force to collect just the right amount and placed it gingerly inside the test tube. This was the perfect dirt sample! Yes you heard right, I said dirt, the brown, clay-textured earth that my parents told me never to play in.  However, in the name of phages, I had to break that rule.

I originally believed that I had collected an extremely fertile dirt sample; following the enrichment of my sample I had yielded around 20 putative plaques.  For those of you reading this who are in my phage hunting class, do you remember the collective sigh you all let out when I presented my original results in the first lab meeting?  Please uptake the oxygen you wasted.  A week later, I lost all of my phage to a deadly contamination; honestly, I probably brought it on myself by coming to class with a sinus infection (insert sigh here). I tried all I could to save my beloved phages from flooding with 5 and 10 mL of phage buffer to CPR (well not really, but I wish I could have).  From there I only had one heartbreaking option, to thrown my children into the trash can of no return and adopt a new phage to isolate as my own.

My new phage, ALS 1aD, named for its original parent, is now on its way to being isolated and sequenced.  I performed a tertiary streak last week of my putative plaques and at the end of last week I began the process of purifying the phage.  As much as I would like to say that this is going smoothly, it is not.  Instead of performing the phage titer assay once and moving on to the next step in the process, I have been forced to perform the assay three, yes THREE, times!  The first time I performed it, the incubator was magically turned off by an unknown being leaving the petri dishes barren and plaqueless.  The second time I performed the assay, the agar on my plates slid, causing some of my plaques to be unable to be counted which would cause error in my calculation.  They say the third time’s a charm so hopefully I will end up with beautiful plates on Friday.  The story of my phage will continue on and it will hopefully progress past the point of the phage olympics.  For now the question remains…phage or not a phage?

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2 Responses to Phage or not a Phage…That is the Question!

  1. emilyjanefisher says:

    Even though your phages ended up being specific to the wrong bacteria, it’s good to know the beach is full of phages. Several other students tried to duplicate your success and found mycobacteriophages there. I think at least 4 people found their phages there after you recommended it!
    The gen. bio. class does a biomes project in which they classify the species of different parts of campus. I told Dr. Pearlman they should add lots and lots of phage to their lists of species.

  2. roboyle1 says:

    Despite the fact that I didn’t find my own phage it feels good to know that I was able to help other people find phages and other bacteria on the beach. Maybe they can rename the beach, the Phage Farm 🙂

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