By Gregory Konar
For those of you who picked up on the title playing on “The Call of the Wild” by Jack London, kudos to you, you must be either a writing sems major or a very well educated other person!
But I digress, already, this post is not about Jack London or literature at all, this is about the wonderful and shockingly populous phage!
This here is a picture of a very sinister looking phage, perhaps he is participating in No Shave November? There are many other phages just like him, thousands of millions of billions of phages exist on the earth at any given moment. In fact, as you read this blog post, you might have typed on a cluster of phages hidden in bacteria on your keyboard! Eww, go clean your keyboard please! All good now? Alrighty, let’s continue on this phage journey!
Phage Hunting is an absolutely amazing class that for me runs Mondays and Wednesdays from 2:30 to 5:00. Yeah, big time commitment for a 2 credit class, but trust me when I say that it is worth it. It is single handedly the greatest class that I have taken this semester! (and no, I am not saying this just to appease the almighty powers that I call Dr. Fisher and Dr. Schildbach)
So, how do we answer the call of the phage? Well first, we had to find a phage!!!
(wise words of advice!)
Soil was our medium of choice for finding the phage, so off I galumphed searching for the perfect patch of dirt, tedious tedious work! It took me a whopping 5 minutes to locate and dig up soil necessary for the lab, and it ended up being pretty darn nice soil!
I tried 2 different methods of coaxing phages from the soil sample: Direct Plating and Enrichment. Direct Plating was like making a pizza, the agar plate was the dough, and then I added the sauce in the form of Mycobacterium smegmatis (the friendly cousin to tuberculosis) and the cheese was the dirt. Enrichment was like steroids, we juiced up the concoction with lots of food and goodies and let it fly. Low and behold, only the enrichment worked, and the pizza had nothing to show for itself.
Once the enrichment provided some putative phage plaques (no, not the bad plaque that forms on your teeth), I went forward and plated and replated and replated the phage 4 times until the plaques were all nice and stable in morphology. Then the titering began…
And as chemistry cat so eloquently puts it, I needed the concentration of phages present on the plates. So the plates were flooded and liquid stocks of the phages were created. This was done once on a small scale to get the MTL and then again on a much larger scale to get the HTL. From the HTL the real fun began!
The HTL unlocked a whole new world of possibilities! From there I did a whole host of things ranging from DNA Preps to Restriction Enzyme digests to Electron Microscopy! The possibilities were endless, and everything was amazing and fun and eye opening! My phage baby had a 48nm head and a 160nm tail, which I thought was pretty darn awesome!
I haven’t ever used an electron microscope before, so this was a new experience for me!
As of the time in which I write this blog, I am busily preparing for the Phage Olympics on top of an RNA presentation, Chem Test, and panic over class registration. So, it will be a fun and great experience on Monday at the Olympics, hopefully my baby will be in tip top shape for that and will bring home the gold! Let’s see if it can answer the call of the phage!
Oh, did I mention that its name is Cell-Fie?
Until next time! Phage out!