When describing how you isolated your phage you mentioned something called “M. Shmegmatis.” You say that you added this to the soil, incubated it, spun it, and filtered it to make lysate. What is the function of M. Shmegmatis and why did you add it to the soil? Can you explain this process a little more?
Yes, M. smegmatis is a strain of bacteria that is non-pathogenic and is closely related to M. tuberculosis. There is no “function” of M. smeg because it is just a bacterial strain, but it is added to the soil in order to isolate the phages. The process of making lysate is exactly as you said, after combining the M. smeg and soil, we incubated the sample, spun it in order to separate the liquid from the soil, and filtered debris out of the liquid so that we could have our pure phage lysate.
Lytic phages have small and clear plaques because they are completely killing the bacterial lawn. Temperate phages have larger and cloudy plaques because they did not kill the bacterial lawn all the way through because they went through the lysogenic life cycle instead of the lytic life cycle.
First we would need to run our phage DNA through a PCR gel with chosen clusters to get an idea of what cluster our phage could possibly be in. After that the phage would need to be sequenced which is when we would find out for sure what cluster our phage is in. After finding out the cluster and subcluster of the phage we would archive it to be used for phage therapy.
When describing how you isolated your phage you mentioned something called “M. Shmegmatis.” You say that you added this to the soil, incubated it, spun it, and filtered it to make lysate. What is the function of M. Shmegmatis and why did you add it to the soil? Can you explain this process a little more?
Yes, M. smegmatis is a strain of bacteria that is non-pathogenic and is closely related to M. tuberculosis. There is no “function” of M. smeg because it is just a bacterial strain, but it is added to the soil in order to isolate the phages. The process of making lysate is exactly as you said, after combining the M. smeg and soil, we incubated the sample, spun it in order to separate the liquid from the soil, and filtered debris out of the liquid so that we could have our pure phage lysate.
What was the hypothesis of your experiment?
We did not have a hypothesis.
Why do lytic phages have small and clear plaques while temperate phages have larger and cloudy plaques?
Lytic phages have small and clear plaques because they are completely killing the bacterial lawn. Temperate phages have larger and cloudy plaques because they did not kill the bacterial lawn all the way through because they went through the lysogenic life cycle instead of the lytic life cycle.
What steps would need to be taken to begin using your phage for phage therapy?
First we would need to run our phage DNA through a PCR gel with chosen clusters to get an idea of what cluster our phage could possibly be in. After that the phage would need to be sequenced which is when we would find out for sure what cluster our phage is in. After finding out the cluster and subcluster of the phage we would archive it to be used for phage therapy.