Phage Issues…

By Ashley Foreman

So after over a month of trying to find and isolate a phage, I’ve definitely run into my fair share of problems. From the very beginning, I should have known this was going to be an uphill battle. I collected three samples, none of which ended up having a phage. I ended up having to “borrow” a plate from a friend and run with that. Then came the next set of problems. It basically goes as such: each time I attempted to streak from a plaque, it resulted in some nasty cases of yellow, smelly contamination. Then, things started to look up as the streaking process continued and I was turning out some beautiful plates. By my third streak, my plates looked like they could be textbook examples, but then the problems returned. I ran into a similar situation with contamination for my first attempt at a titer assay and had to do several trials of serial dilutions until I got something usable. Then, I attempted to get a medium titer lysate but it was contaminated so I tried again, and it turned out to be, yet again, contaminated. So, I ended up filtering the flooded plate to try to avoid any bacteria. Finally, I got an MTL! Unfortunately, after I calculated my max web and was prepped to start plating a max web, I got laryngitis and had to miss class. When I returned, my MTL PB sample was gone!! So, I had to go back to the closest thing I had which was my 10-7 sample and serially dilute that to try to get a new MTL. I just got the results from that, most of which were contaminated (go figure) but my 10-3 sample ended up being close enough to a max web for me to start plating 6 samples of it. This week, I’m going to have to log some extra hours in the lab to finally try to get a high titer lysate in order to try to get even minutely caught up. Hopefully, contamination won’t play a role in this process. Wish me luck! Image

My “max web!”

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