By Madison Wahl
Entering into the magical world of bacteriophages is an exciting and promising time for any young researcher. Researchers are desperate to make their mark on the world, and finding, isolating, and taming the perfect phage can be their way to do just that. And this will most certainly be a wonderful and eventful time. Raising a phage will unleash a world of dedication, commitment, and an unprecedented form of love from the researcher.
However, contrary to the Hallmark hysteria, this will not be a time only of giggles and smiles and raspberry kisses. It will be perhaps one of the most trying times for the researcher as he/she learns how to balance being a researcher and properly taming his/her phage, and dealing with all the challenges and fits put out by the one they so love. So there are a few things you should keep in mind that will help make the transition a little easier as you enter into this electrifying new experience.
For starters, a mutual relationship of trust and dedication is essential to a successful phage. From the minute you begin the direct plating, you need to be confident in your sample’s ability to produce a solid and healthy phage. And, once you begin isolating your phage, you must be confident in your phage’s ability to be isolated, provide a high enough titer, et cetera. This confidence may be difficult at times, particularly if your phage does not want to isolate or will not produce a high-enough DNA count. But your job as the researcher and as its founder is to keep trying. Keep pursuing a stronger, healthier phage, and in time your phage will deliver.
Another key to remember is that socializing your phage is NOT recommended for a successful isolation and taming process. It may be tempting to schedule play sessions with your lab partner’s phage, or to use hand-me-down streaking sticks or pipette tips from another phage, but this will only lead to trouble. Your phage will thrive–and your relationship will grow even stronger– if you keep your phage away from any possible contact with any other phages. It might feel like you have to be the bad guy, but again that is your job as the researcher. A pure strand of isolated phage is necessary for a proper titer and DNA concentration.
There will be days when you have had enough. When the lab bench is a mess and the bunsen burner/lab coat combination is causing you to sweat bullets and there are no centrifuges open and your phage will not cooperate with the DNA prep you are trying to do and you are running on two hours of sleep from staying up to finish your math homework because you were nursing your phage to health the previous day and you are ready to pull out your hair and sit in a puddle of your own tears mixed with the 80% isopropyl your phage caused you to spill on the floor. It’s okay to have these days. We all do. It’s okay to be frustrated and upset. It’s okay to take it easy that day and save DNA prep for tomorrow. And it’s always okay to call your mom, because even if she never raised a phage, she somehow seems to have all the answers. But it’s not okay to stop trying–to throw in the towel and abandon your phage. What you need to do is take these days in stride. Take a deep breath and remember all the wonderful days of streaking you had with your phage.. all the smiles as he repeatedly evoked as you walked into lab to greet him and saw that his plaques were superb. Remember why you chose to raise a phage, pick yourself up, dust yourself off, and get back to work. After all, he needs you and it will get better, I promise.
And lastly, don’t expect others to see your phage the way you do. Where others see a heap of dirt, you see your un-isolated phage. Where others see small lytic plaques, you see your phage isolating remarkably, light years ahead of all the rest. Where others see a seemingly average DNA concentration, you see the days of sweat and tears that brought that concentration up from 5 to 80. When others see a phage with too short of a tail and too big of a head, you see the unique characteristics of a phage that has created such beautiful plaques. Do not let the world harden you. Continue you look at your phage with the same sparkle in your eyes the first day you found, because you really are his one and only.
Cruz and I have gone through many of these trying times together. His plaques were worrisome at first, given the change in size, but we figured out together that that’s just who he was. His DNA concentration was initially very low, but we worked through that together to give a very impressive number in the end (even though one column broke and we were only able to use half of the DNA). His max-web plates were difficult to find and caused many days of heartache, but that, too, we solved. It really is a labor of love when you choose to harvest and isolate a phage, and if you are doing it for the right reasons it will certainly be one of the most rewarding experiences you will have as a researcher.