What could be a more effective mixing technique you could use in the future so that the result that you got in your lane 2 of your gel doesn’t happen again?
The error made in mixing our DNA during the quality control portion of the experiment is so trivial and obvious that it’s honestly disappointing. We know that lane one yielded results and both solutions contained DNA from the same isolation procedure, so we know our DNA is not the issue. When creating the solution used to put into the wells during gel electrophoresis, the amount of liquid used is so small, especially with regard to DNA, that it can be hard to properly eject the solution from the pipette tip. We added our DNA to the centrifuge tube first using only 2µL, which likely got stuck on the side of the tube. Adding the water and loading dye first, and then adding DNA directly into the liquid, likely would have resulted in a more homogenous solution. Additionally, pipetting the solution up and down to mix is a must.
Based off the microscopic image you got of your phage, can you predict whether the phage is lytic/lysogenic, or predict what cluster is may be a part of?
Based on the morphology of our phage being a temperate phage, meaning it has the option to undergo both the lytic/lysogenic lifecycles, it is likely that both are occurring. When looking at our plate we can see that the plaques are cloudy meaning that most of the cells are lysed (roughly 80%) and some undergo lysogeny (roughly 20%) becoming a prophage. if the plaques were fully clear we would know that the phage is purely lytic, that is not the case however
With regard to cluster, we predicted either cluster A or B as the EM image appears to fit into A or B cluster, which are some of the most common. This is not a reliable result, but a prediction/hypothesis. We would need to perform PCR in order to concretely determine the cluster. Our quality control results did not yield the results we wanted due to error in DNA isolation, so we would need to re-do that experiment as well as restriction digest and PCR to get a reliable cluster assignment.
Yes! For bacterial infections occurring inside the body, the phage treatment would be administered intravenously, using the titer calculation to determine the correct dosage of phage. If an infection were to occur on the skin, the treatment would likely be administered topically.
curesymposium.org 
What could be a more effective mixing technique you could use in the future so that the result that you got in your lane 2 of your gel doesn’t happen again?
LikeLike
The error made in mixing our DNA during the quality control portion of the experiment is so trivial and obvious that it’s honestly disappointing. We know that lane one yielded results and both solutions contained DNA from the same isolation procedure, so we know our DNA is not the issue. When creating the solution used to put into the wells during gel electrophoresis, the amount of liquid used is so small, especially with regard to DNA, that it can be hard to properly eject the solution from the pipette tip. We added our DNA to the centrifuge tube first using only 2µL, which likely got stuck on the side of the tube. Adding the water and loading dye first, and then adding DNA directly into the liquid, likely would have resulted in a more homogenous solution. Additionally, pipetting the solution up and down to mix is a must.
LikeLike
Based off the microscopic image you got of your phage, can you predict whether the phage is lytic/lysogenic, or predict what cluster is may be a part of?
LikeLike
Based on the morphology of our phage being a temperate phage, meaning it has the option to undergo both the lytic/lysogenic lifecycles, it is likely that both are occurring. When looking at our plate we can see that the plaques are cloudy meaning that most of the cells are lysed (roughly 80%) and some undergo lysogeny (roughly 20%) becoming a prophage. if the plaques were fully clear we would know that the phage is purely lytic, that is not the case however
With regard to cluster, we predicted either cluster A or B as the EM image appears to fit into A or B cluster, which are some of the most common. This is not a reliable result, but a prediction/hypothesis. We would need to perform PCR in order to concretely determine the cluster. Our quality control results did not yield the results we wanted due to error in DNA isolation, so we would need to re-do that experiment as well as restriction digest and PCR to get a reliable cluster assignment.
LikeLike
If this were eventually used in treatment would it be administered intravenously?
LikeLike
Yes! For bacterial infections occurring inside the body, the phage treatment would be administered intravenously, using the titer calculation to determine the correct dosage of phage. If an infection were to occur on the skin, the treatment would likely be administered topically.
LikeLike