Cluster primers are PCR primers that are specific to a certain cluster of phage. When you add phage DNA to PCR mix along with specific primers, you are able to tell which cluster your phage belongs to. I used L and B in this case, so when I check my gel, I will be looking for DNA bands present in the L and B lanes, which indicates that my phage DNA has bound to the primers, thus indicating which cluster it belongs in.
M. smeg bacteria are non-pathogenic to humans and animals, but some phages that can infect them also happen to be able to infect bacterium that are pathogenic to humans. Getting a base off of M. smeg can help to find phages that can be used in therapeutic measures
Phage morphology can fall under S type or M type, myoviridae. The biggest differences are that S type have long, non-contractile tails and M-type have Short, contractile tails. S-type phage can have both circular and hotdog shaped heads, and M-type are restricted to spherical heads.
Most phages identified are S-type
We essentially isolated a phage from a random dirt sample found on campus in order to determine its morphology. Certain types of phages can be useful in different areas of medicine. You can use lytic phages to fight bacterial infection, like in the case of the girl with CF. With a temperate phage, there is a thing called phage display, which works as an antivenom for snakebites. Ideally I would love to see if I could implicate my phage in such display to increase the range of antivenoms.
We didn’t have a set number of enzymes that “should” cut the DNA, because we performed the digest as a method of estimating which cluster was the best match. However, we did expect at least one enzyme to cut, though to be honest I was surprised that more enzymes didn’t cut it (especially EcoRI). I am used to seeing EcoRI cut everything, so I was surprised that this enzyme did not cut. If I had the time, I would run the digest again and maybe even perform the DNA isolation a second time.
Can you please explain what cluster primers are?
Cluster primers are PCR primers that are specific to a certain cluster of phage. When you add phage DNA to PCR mix along with specific primers, you are able to tell which cluster your phage belongs to. I used L and B in this case, so when I check my gel, I will be looking for DNA bands present in the L and B lanes, which indicates that my phage DNA has bound to the primers, thus indicating which cluster it belongs in.
Why were M. smeg bacteria used?
M. smeg bacteria are non-pathogenic to humans and animals, but some phages that can infect them also happen to be able to infect bacterium that are pathogenic to humans. Getting a base off of M. smeg can help to find phages that can be used in therapeutic measures
Can you explain spirvirdae compared to what other kinds are.
Phage morphology can fall under S type or M type, myoviridae. The biggest differences are that S type have long, non-contractile tails and M-type have Short, contractile tails. S-type phage can have both circular and hotdog shaped heads, and M-type are restricted to spherical heads.
Most phages identified are S-type
Could you explain the purpose of this experiment a little bit more?
We essentially isolated a phage from a random dirt sample found on campus in order to determine its morphology. Certain types of phages can be useful in different areas of medicine. You can use lytic phages to fight bacterial infection, like in the case of the girl with CF. With a temperate phage, there is a thing called phage display, which works as an antivenom for snakebites. Ideally I would love to see if I could implicate my phage in such display to increase the range of antivenoms.
How many enzymes were you expecting to be successful to cut the DNA?
We didn’t have a set number of enzymes that “should” cut the DNA, because we performed the digest as a method of estimating which cluster was the best match. However, we did expect at least one enzyme to cut, though to be honest I was surprised that more enzymes didn’t cut it (especially EcoRI). I am used to seeing EcoRI cut everything, so I was surprised that this enzyme did not cut. If I had the time, I would run the digest again and maybe even perform the DNA isolation a second time.