We made 4 1:10 serial dilutions of our phage lysate because this gave us a selection to see which concentration produced the clearest and most dilute plaques to use for our web pattern plate. We used 10^-1, 10^-2, 10^-3, and 10^-4 for this step.
Hi Narges! We concluded that our DNA was high quantity due to its bright fluorescent color produced from the gel electrophoresis. If it were very faint or non-visible, it could be due to a low amount of DNA or a total absence of DNA.
What is phage purification as mentioned in your presentation and how did you determine whether or not your phage in specific was able to move on to this stage?
Phage purification is used to isolate one phage from a plaque assay. This is to ensure only one phage is being analyzed throughout the process and to eliminate contamination. We determined that our phage was purified enough as we performed two separate rounds of phage purification to ensure only one phage would be left. Luckily, we also did not observe any differences in the phage morphology present on our plaque assays, so we were sure we were only dealing with a single phage.
Hi Kayla! Phage purification is the process of isolating one phage for analysis. We determined our phage was ready to move onto phage purification as it had produced a clear and dilute plaque assay from the phage lysate.
Sadly, no, we don’t have any guesses as to what enzymes would have been able to cut the DNA. Though, most phages at Boulder are found to be in cluster A, so that could be something to keep in mind as we look into in the future if restriction digest is able to be ran on the phage.
How did you decide what concentrations to use for serial dilutions?
We made 4 1:10 serial dilutions of our phage lysate because this gave us a selection to see which concentration produced the clearest and most dilute plaques to use for our web pattern plate. We used 10^-1, 10^-2, 10^-3, and 10^-4 for this step.
Hi Elizabeth, Why did you conclude that your gel was high quantity?
Hi Narges! We concluded that our DNA was high quantity due to its bright fluorescent color produced from the gel electrophoresis. If it were very faint or non-visible, it could be due to a low amount of DNA or a total absence of DNA.
What is phage purification as mentioned in your presentation and how did you determine whether or not your phage in specific was able to move on to this stage?
Phage purification is used to isolate one phage from a plaque assay. This is to ensure only one phage is being analyzed throughout the process and to eliminate contamination. We determined that our phage was purified enough as we performed two separate rounds of phage purification to ensure only one phage would be left. Luckily, we also did not observe any differences in the phage morphology present on our plaque assays, so we were sure we were only dealing with a single phage.
Hi Kayla! Phage purification is the process of isolating one phage for analysis. We determined our phage was ready to move onto phage purification as it had produced a clear and dilute plaque assay from the phage lysate.
Although you were unable to run restriction digest on your sample, do you have any predictions about which enzymes would cut the DNA?
Sadly, no, we don’t have any guesses as to what enzymes would have been able to cut the DNA. Though, most phages at Boulder are found to be in cluster A, so that could be something to keep in mind as we look into in the future if restriction digest is able to be ran on the phage.