8 thoughts on “P40 – Miller

  1. Great work! I’m curious about if your procedure would have looked any different if you had found the phage to not be temperate?

    1. If it had been found that the phage was not temperate, we would have known the phage was lytic. Our procedure would be unaffected as the goal during the lab is to just characterize the phage we had isolated and has the same protocols despite the lifecycle. We would know throughout the procedure though that our phage was undergoing the lytic lifecycle and completely lysing the bacteria.

  2. I really enjoyed your presentation! Are there any specific conditions that you think this phage could effectively treat?

    1. At this point in time, the phage would not be able to effectively treat any conditions. We were not able to complete PCR so were only able to create a hypothesis on what subcluster the phage was part of through restriction digest. Knowing the cluster is important to understanding what the phage could be used for. Also, to be used to treat an infection our phage would need to be made lytic through the removal of the integrase and repressor proteins. If both were done, we would be able to learn what the phage could be used for effectively.

  3. Good job on your presentation! I’m curious to know where the dirt sample used to isolate the phage was collected from?

    1. The dirt sample was collected outside Folsom field within the trees and bushes surrounding the stadium. The soil was dug out from under wood chips. This was then enriched through being mixed with LB Media and M. smeg which was then purified through dilutions so were able to isolate the phage.

  4. What would performing a PCR to confirm the phage’s cluster look like? Would you have primers specific to the K cluster to see if they are amplified?

    1. PCR would involve amplifying DNA sequences to confirm the cluster. We would use primers of the K cluster as well as N, because it had the next highest similarity with the enzyme tool. Our phage DNA would then be combined with these primers and a PCR mixture, and run through a 0.8% agarose gel as well as positive and negative controls. Once amplified, we could determine if the primers had been successful.

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