Our best guess is improper loading of the gel. The enzyme in that column is Haelll, so we were expecting a multitude of cuts throughout the entire column.
On our original plate from the plaque assay experiment, we actually had both temperate and lytic phages. We just happened to isolate a lytic phage by streaking and performing serial dilutions from a lytic plaque.
We wanted to the count the cuts, so we could put them in Phage Enzyme Tools 2.0. From there, we could hypothesize what cluster it would be in. Then, we would perform a PCR (with the relevant primers) to confirm or disprove our hypothesis.
I received the image in green. Prior to posting this, I edited the photo to black and white; however, the image quality was absolute garbage, so I decided to nix it in favor of the green.
Absolutely. You can manipulate temperature, use different solvents, use enzymes, physical stimulation, etc. In the context of this experiment, we might not want to deliberately weaken the cell wall, since we are only characterizing a novel phage. However, if we want examine how efficiently our phage inserts its genetic material, we might play around with the aforementioned methods.
Why is your sixth well in your gel electrophoresis figure is mostly blank and then has a bulk of brighter bands?
What is the reason for using lytic bacteriophage instead of lysogenic?
Our best guess is improper loading of the gel. The enzyme in that column is Haelll, so we were expecting a multitude of cuts throughout the entire column.
On our original plate from the plaque assay experiment, we actually had both temperate and lytic phages. We just happened to isolate a lytic phage by streaking and performing serial dilutions from a lytic plaque.
I got slightly confused on what the gel electrophoresis was for? Also, as someone who has done them before, why is it green?
We wanted to the count the cuts, so we could put them in Phage Enzyme Tools 2.0. From there, we could hypothesize what cluster it would be in. Then, we would perform a PCR (with the relevant primers) to confirm or disprove our hypothesis.
I received the image in green. Prior to posting this, I edited the photo to black and white; however, the image quality was absolute garbage, so I decided to nix it in favor of the green.
Is there a way to weaken the cell wall artifically so the phage can break through?
Absolutely. You can manipulate temperature, use different solvents, use enzymes, physical stimulation, etc. In the context of this experiment, we might not want to deliberately weaken the cell wall, since we are only characterizing a novel phage. However, if we want examine how efficiently our phage inserts its genetic material, we might play around with the aforementioned methods.