by Samantha Siow
It’s been a long journey.
There have been too many times when my partner and I have asked ourselves, “How much longer? Where are the phages? Is there something wrong with us? Do we smell bad? Is it our pipetting? Is it our terribly fashionable purple and pink disposable lab coats? Where them phages at?”
But no longer.
I would go into detail about the emotional turmoil that we found ourselves knee-deep in, the endless pits of despair, the times we found ourselves staring at a building pile of blank plates and felt the crushing disappointment of the failure of another collection of ‘phage-filled’ soil samples.
It’s okay – times are better now. We’ve moved on.
Thinking back, there was no such thing as a failure in Phage Hunting. With every direct plating or enrichment, we became better. Our skill was rivaled across the land (or at least on our own bench). We pipetted at incredible speeds. Not a bubble was seen on our plates. Yet, not a phage was seen as well. But with every negative plate, we improved. We knew the time would come. The phages were out there. Waiting.
And it did. It was a magical day. Dr Schildbach’s compost sample, enriched to make it as wonderfully nasty as possible, yielded positive results. It was our second enrichment after what felt like a billion negative direct platings, and how we celebrated.
But it did not stop there. The hands of fate (or should I say Phate? Haha.) had more in store for us. Hungry for a phage to call our own, Peter (my awesome lab partner) and I scoured the lands and found ourselves on a hill near Bloomberg on a beautiful day. Crouched down and poking at the ground with our technologically advanced plastic utensils, we collected our samples. They weren’t unique or spectacular. It was soft, moist soil like any other and I did not expect much. I named it “The Boss” in hopes of something promising – Dr. Schildbach’s compost phage had been titled “Boss” and we figured that that must be the key to all phages – and left it to enrich. We must have been right.
Since the Original Boss’s founding, we have been having positive results from samples all around. Although Original Boss is not our own, he will always be special to us. We’re proud parents of the phage that is most ‘phar’ ahead, having achieved a beautiful, awesomely psychedelic web (photos to be added later) from our first titer in dilution 10^-3 after 3 perfect streaks, flooded with phage buffer, and extracted. Our spot test to some M. smeg yielded unsatisfactory results from our different dilutions, with less definition between plaques then we would like, but we are currently in the process of plating each phage dilution from 10-6 to 10-10 to separate plates in hope of some sweet plaques. The Mysterious Case of the Incubator and The Unsolidifying Agar have so far made our efforts fail, but this Friday should show some promise in bringing us one step closer calculating the dilution needed to achieve the max web. The Boss is also well on his way, and will be ready for titer by this Friday. Its plaques are much cloudier, more translucent and fuzzier around the edges that Boss’s clear, well-defined ones. I like to think of him as the quiet genius kid while Boss is the one who would steal his lunch money. But they’re both equally awesome. No favorites.
Just some tips I’ve learned from my many failures:
- When pipetting up the TA after mixing with the M. smeg, DO NOT pipette all the solution back up. Pipette back up slowly and leave just the tiniest volume left in the test tube. This will ensure no bubbles are left in the TA when deposited back onto the plate.
- Work fast! If you see any bubbles on your plate, pipette it back up gently and slowly before the TA has any time to set. They can look like fake plaques that only prove to crush your dreams when you find yourself staring at a negative streak test. It’s okay. We’ve all been there before.
- Slow and steady CAN win the race. Our lack of phages allowed us to escape the Great Contamination of ’11. None of our phages were lost… simply because we had none.
- HIFIVES are very important. Catch a bubble right at the end of your pipette? HIFIVE. Keep the motivation up when phages elude you!
- Lastly, don’t lose hope! Your phage is out there somewhere. They’re just making sure you’re good enough. It’s all an initiation process.
To all the future phage hunters out there,
MAY THE PHAGE BE WITH YOU.