What type of future testing is needed to determine the cluster of the particular phage and how will your limitations of this research impact your approach to future testing?
Hi Kayla, thank you for your question. For determining the cluster of our phage we would need to first get a successful quality control gel to confirm that the DNA is both high enough quantity and quality. Once that has been achieved we would do a restriction digest on the DNA and match our band pattern to other clusters band patterns.
Hi Madison, thank you so much for your comment! A 0.8% agarose gel is standard for the size of DNA fragment that we used. Depending on the agarose concentration in your gel, DNA will either travel faster or slower through the gel. This one is also the most efficient concentration for the quality controls that we are running.
We think that the major factor that contributed to our unsuccessful quality control gel was the quality of the DNA that we were running on it. Since we ran the control gel multiple times with multiple different protocols, we determined that the protocol was not the problem. Our high titer lysate(which contained our phage DNA) was likely not high quality enough to have a successful quality control gel. However, that’s what those gels are for!
Generally, the shape of the virus is not related to whether the virus is lytic or lysogenic. Lytic and lysogenic virus classifications refer to the lifecycle and behavior of the phage while shape is juts about visual appearance.
Great Presentation Emma! You had a really clear explanation for the big research, and walking us through the process your team did to characterize your phage!
What type of future testing is needed to determine the cluster of the particular phage and how will your limitations of this research impact your approach to future testing?
Hi Kayla, thank you for your question. For determining the cluster of our phage we would need to first get a successful quality control gel to confirm that the DNA is both high enough quantity and quality. Once that has been achieved we would do a restriction digest on the DNA and match our band pattern to other clusters band patterns.
Why was a 0.8% agarose gel chosen over a different percentage gel? Great job on your presentation!
Hi Madison, thank you so much for your comment! A 0.8% agarose gel is standard for the size of DNA fragment that we used. Depending on the agarose concentration in your gel, DNA will either travel faster or slower through the gel. This one is also the most efficient concentration for the quality controls that we are running.
What do you think were some of the major factors that lead to a faulty quality control gel?
We think that the major factor that contributed to our unsuccessful quality control gel was the quality of the DNA that we were running on it. Since we ran the control gel multiple times with multiple different protocols, we determined that the protocol was not the problem. Our high titer lysate(which contained our phage DNA) was likely not high quality enough to have a successful quality control gel. However, that’s what those gels are for!
Does the shape of the virus, such as it being the long or short one, effect whether the virus will be lytic or lysogenic?
Generally, the shape of the virus is not related to whether the virus is lytic or lysogenic. Lytic and lysogenic virus classifications refer to the lifecycle and behavior of the phage while shape is juts about visual appearance.
Great Presentation Emma! You had a really clear explanation for the big research, and walking us through the process your team did to characterize your phage!