The Quest for Phage

By Allen Chen

When I first received an email about homework for phage lab, I thought it was funny that the very first assignment of my college career was to go dig up some dirt. I remember putting it off until the day before the first day of lab, which in retrospect was a bad idea because it ended up raining. As punishment for my poor work ethic, I had to dig up my unpleasantly wet soil sample with my bare hand. Luckily for me, it was dark, so at the very least I can say that nobody saw me.

The first few days in lab were difficult. The procedures seemed unnecessarily complicated and lengthy. I remember having to redo my first attempt at a serial dilution because I had forgotten to add the phage onto the plates I was making, resulting in plates that only had top agar on them. On top of that, my plates were upside-down, so the top agar was sitting by itself on an empty lid.

Luckily, I got better and faster at these techniques, and I began to enjoy the project. Finding plaques on my enrichment sample was particularly exciting. Back in high school, I was generally horrible at labs, and seeing the phage on the enrichment plates meant I had actually done something right, which was mildly surprising. I had two different phage morphologies. The first was large and had a fuzzy outline; the second was smaller with a more clearly defined outline. I was definitely a proud parent.

Once I had obtained phages, I began the long process of streaking my plates, and eventually, I was able to purify a single phage population. Currently, I am in the process of creating a webbed plate, or one that is completely covered in a thin layer of phage. This will allow me to make a high-titer lysate (HTL). Right now, I have no idea what that is, but I look forward to learning more about it and the procedure.

So far, I’ve had a blast in this course. I never imagined I’d be able to do something so fun and hands-on during my first semester in college, and it’s been a huge step up from high school. I can’t wait to see how the rest of the year turns out!

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4 Responses to The Quest for Phage

  1. Pingback: The Quest for Phage | jhublogs

  2. epresse1 says:

    I too struggled with getting pure phage from streaking. It took me seven rounds before I was confident I had pure phage! I also initially had two different phages, a phage that produced larger, turbid plaques and a phage that produced really tiny, clear plaques. I ended up choosing to continue with my small phage.

  3. yooashley says:

    The entire process had a great deal of learning. I can completely relate with the fact that the first few days in lab put me in a whirlwind of confusion. Despite that, I think I speak for all of us when I say that we really learned to use our own intuition to problem solve and work with the most frustrating of phages. Hope the rest of your process went smoothly!

  4. mchen78 says:

    You are not alone. I too, had made some terrible mistakes along the way which delayed my process, such as not vortexing the phage buffer mixture properly before performing the next step of the dilution series. This just shows that a lab procedure isn’t ground in stone, and must be tailored according to the situation. I hope the rest of your experiment turned out well!

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