View from Up Ahead

By Sumukh Shetty

What is phage hunting? Is it worth my time at the tail end of a six hour day or is sleep more important? As the 7am registration deadline for selecting classes on ISIS approached, these questions bombarded my sleep-deprived mind. It was in my class cart only because my adviser told me to take it to fulfill credit requirements. Eventually I decided to take the risk and registered for the class. When my friends heard about it, they kept questioning me about the class. I joked with them that it was a hybrid between what Elmer Fudd did and what Bill Nye did. Only once lab started did I realize that the micropipettor would be the weapon of my choice and my soil sample would be my enemy. One does not simply hunt for phages.

SMS1

Fast forward a month and phage hunting is now my daily respite on Mondays and Wednesdays after 5 hours of class. I’ve performed so many serial dilutions and plated so many smeg plates that I can do them in my sleep. SMS2

Eventually I got my high-titer lysate and was done plating or so I thought. Following the HTL, I decided to view my phage under a electron microscope. With the help of Mr. McCaffery at the IIC, I finally got to see the phage that I had spent the last month streaking and plating. And it did not disappoint. Originally all I saw were tails and heads that had been separated along with some organelles. At a different location, however, we found a group of perfect phages. They were a type of siphoviridae with heads roughly 95nm and tails that were 275 nm long. I was so relieved to have such a good result after all the work.

SMS EM

Following the electron microscopy, I had to isolate and purify the DNA of my phage. After precipitating the phage particles and using some elbow grease to isolate the DNA, I finally had a small amount of my DNA. But what’s the point of having some DNA if you don’t electrophorese it in a gel? After a process that seemed more boring than watching paint dry, the gel was finally done. Not only did it indicate that my DNA was properly cut up, but also it showed that I didn’t mess up doing the gel like I have in the past. Success!!! I can only hope this continues because now as I type this, my leftover restricted DNA is hopefully being spliced into a plasmid for sequencing.

SMS3

Whatever doubts I had when registering have completely disappeared. Instead I can’t wait till it’s 2:30 on Monday or Wednesday. Each day is a new adventure in the world of phages.

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One Response to View from Up Ahead

  1. Pingback: View from Up Ahead | jhublogs

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