My phage has the wonderful name of Lorge. This name is very special because it offers information about the phage right away. I also chose this name because the first word that Tracy said when she saw my phage was, “Wow, it’s huge!”
To get this wonderful phage I collected the soil sample from the flower pots across the street outside of Bessey Hall. I got the sample at the start of class on September 11, 2017. It was a clear day that was 60 degrees, with a depth of about 4cm below the surface of the soil. After I collected my sample I filled the conical tube with enrichment broth to the 20-millimeter line. The solution was then filtered through a coffee filter, to get the larger particulates out, and then the solution was filtered through a …show more content…
The first time I did a plaque assay my phage was at a concentration of 7.2*10^6 pfu/ml. The second time I did a plaque assay my concentration was fairly similar, a little lower, it was 2.8*10^6 pfu/ml. From this second round of purification, I picked a relatively isolated plaque and used that to do a serial dilution, from this dilution I could do my first spot titer test. This test would allow me to collect my small volume lysate, and calculate the concentration of my small volume lysate which happened to be roughly 3.3*10^10. This spot titer test gave me the opportunity to calculate what dilution I should do in order to consistently make webbed plates. I made my first webbed plates and flooded them, after letting my webbed plates sit for a little over an hour I was able to collect the solution from the surface of the plate. This solution became my large volume lysate. I repeated the steps of making webbed plates until my large volume lysate was at 9 ml, with a concentration of 9.0*10^9 …show more content…
As shown by the gel electrophoresis my DNA is very clean because, as shown by the picture my uncut DNA has very crisp and clean band in the gel. I wanted this one clean band to show that my DNA is not pre-cut in any way and is pure. The gel also shows that my DNA was cut by Ban II Cla, Eco, Hae III, and Sal enzymes. The gel shows that in these columns the DNA became spread out, and looks more like a smear on the gel, rather than one crisp band like the uncut
To get this wonderful phage I collected the soil sample from the flower pots across the street outside of Bessey Hall. I got the sample at the start of class on September 11, 2017. It was a clear day that was 60 degrees, with a depth of about 4cm below the surface of the soil. After I collected my sample I filled the conical tube with enrichment broth to the 20-millimeter line. The solution was then filtered through a coffee filter, to get the larger particulates out, and then the solution was filtered through a …show more content…
The first time I did a plaque assay my phage was at a concentration of 7.2*10^6 pfu/ml. The second time I did a plaque assay my concentration was fairly similar, a little lower, it was 2.8*10^6 pfu/ml. From this second round of purification, I picked a relatively isolated plaque and used that to do a serial dilution, from this dilution I could do my first spot titer test. This test would allow me to collect my small volume lysate, and calculate the concentration of my small volume lysate which happened to be roughly 3.3*10^10. This spot titer test gave me the opportunity to calculate what dilution I should do in order to consistently make webbed plates. I made my first webbed plates and flooded them, after letting my webbed plates sit for a little over an hour I was able to collect the solution from the surface of the plate. This solution became my large volume lysate. I repeated the steps of making webbed plates until my large volume lysate was at 9 ml, with a concentration of 9.0*10^9 …show more content…
As shown by the gel electrophoresis my DNA is very clean because, as shown by the picture my uncut DNA has very crisp and clean band in the gel. I wanted this one clean band to show that my DNA is not pre-cut in any way and is pure. The gel also shows that my DNA was cut by Ban II Cla, Eco, Hae III, and Sal enzymes. The gel shows that in these columns the DNA became spread out, and looks more like a smear on the gel, rather than one crisp band like the uncut