Great question! Agarose quality control gel is used to see if the DNA which we isolated was of good quantity as well as good quality by comparing it to a DNA ladder.
Can you summarize your methods more cohesively? I do not understand how you went about this experiment. I don’t really know most biological methods besides those from Drug Discovery.
We adopted a phage from another group, therefore it was already enriched and was taken from a spot test. Our experiments started at purification, which is a series of dilutions with phage buffer to try and isolate a single phage in the phage lysate. (which is what was created using a phage from the spot test) We then created a web pattern plate which we used to calculate the titer of the lysate, this is determining how strong the phage that we have isolated was. We then isolated DNA using a DNA wizard column and centrifuging it to push all of the possible DNA through it. We then ran a quality control gel using the DNA which we had isolated, which helped us see if the DNA was of good quality and quantity.
In the lytic lifecycle there are sharp clear cut edges as it completely lyses (kills) the cell, therefore the edges are clear, while temperate phages show a cloudy halo edge because of the growth of the daughter bacteria cells which survive. While others get forced into the lytic lifecycle. This is why it was temperate, because the center was clear where the phages were forced into the lytic lifecycle and the ones on the outside were going through the lysogenic lifecycle.
We adopted this phage from another group, so they got the dirt sample by a bridge between two buildings, with snow on the ground, and a temperature of 27 degrees F.
What exactly is the agarose quality control gel used for?
Great question! Agarose quality control gel is used to see if the DNA which we isolated was of good quantity as well as good quality by comparing it to a DNA ladder.
Can you summarize your methods more cohesively? I do not understand how you went about this experiment. I don’t really know most biological methods besides those from Drug Discovery.
We adopted a phage from another group, therefore it was already enriched and was taken from a spot test. Our experiments started at purification, which is a series of dilutions with phage buffer to try and isolate a single phage in the phage lysate. (which is what was created using a phage from the spot test) We then created a web pattern plate which we used to calculate the titer of the lysate, this is determining how strong the phage that we have isolated was. We then isolated DNA using a DNA wizard column and centrifuging it to push all of the possible DNA through it. We then ran a quality control gel using the DNA which we had isolated, which helped us see if the DNA was of good quality and quantity.
Great presentation! Will you explain why the cloudy, halo edge made you conclude that it was temperate?
In the lytic lifecycle there are sharp clear cut edges as it completely lyses (kills) the cell, therefore the edges are clear, while temperate phages show a cloudy halo edge because of the growth of the daughter bacteria cells which survive. While others get forced into the lytic lifecycle. This is why it was temperate, because the center was clear where the phages were forced into the lytic lifecycle and the ones on the outside were going through the lysogenic lifecycle.
Where did you get your dirt sample from on campus?
Also, what name did y’all give to the phage?
PaolaToscani is the name of our phage
We adopted this phage from another group, so they got the dirt sample by a bridge between two buildings, with snow on the ground, and a temperature of 27 degrees F.