Why Phage Hunting is Even More Complicated Than it Sounds

By Michael Xu

Most people’s preconceptions about this class is accurate, I think. Two sessions, five hours a week, we spend our time in the Macaulay 202 lab room in lab coats and gloves and with bacteria-infested culture plates. We transfer liquids with fancy techniques and we pipette culture with fancy equipment. In a (mostly) independent manner, we each try to find our own special and unique strains of bacteriophages. Sounds like we know what we’re doing, right?

Well, we don’t. Take the first day of class, for example. Most of us had never even lit a bunsen burner, used a micropipette, or made a bacteria culture in our lives and we had to make a direct sample of some soil, note: ON THE FIRST DAY OF CLASS.

Whenever we get the hang of something, there’s just a new and “exciting” procedure to learn. When we thought we finally understood direct plating, we had to learn how to do enrichments. And then streaking. And then titer assaying. And then harvesting lysates. It’s a never ending cycle of confusion.

The most confusing part for me by far was when I was trying to isolate a single type of phage on my plates, and their morphologies kept changing. During my streaking procedure, I created several “generations” of phage as I was trying to isolate a single type of phage within a single colony in successive platings. I started out with one plate with two plaque morphologies, a “small” (S) one and a “large” (L) one. I streaked from each type of plaque to obtain two separate plates, each supposedly with phage from the plaque that I streaked from. The result, strangely enough, was that BOTH plates still had BOTH morphologies of plaque. So then I streaked all four of the resulting phage, and then ended up streaking the subsequent results as well. What was already a daunting and confusing procedure simply became a complete headache when I ended up with a total of around 15 plates, on which I labelled cryptic tags such as LLLL, SLLS, LSS, SSS, and SSL (it was the only way for me to keep track of each plaque’s “family tree”).

If this blog post was unclear and confusing: Good. Because that’s what we’re dealing with every single time we walk in the lab. And, frankly speaking, it’s also why we enjoy every single minute of it.

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