Why do you believe your phage exhibited both lytic and lysogenic lifecycles? Is it possible the environment these plates were stored in was conducive enough to have the lysogenic phages enter the lytic lifecycle some of the time (because you mentioned the lysogenic phages could enter the lytic lifecycle under certain environmental conditions)? Or could it be something else?
Mainly, due to the two different morphologies present in our plaque assays/streaks, we believed that our one phage exhibited both lifecycles – or, we thought there was two phages present. You make a great point though that I didn’t consider! It is definitely possible that the environment in which our phage was stored caused that shift, and is really something interesting to study/ponder. If not, I would assume that we failed to isolate the other phage in EM imaging and there were in fact actually two phages. Thank you for your questions!
Those two lanes are isolated DNA from a phage streak. We wanted to conduct PCR and other characterization tests but due to timing, were unable to run these experiments. The lanes simply represent the presence of our phage’s DNA and confirm to us that we didn’t have some odd contamination (even though the plaques show this as well). Thank you for questions!
I really enjoyed your poster board presentation! I showed you had a lot of knowledge of both your research as well as background into the field. I did have one question regarding your figures and results section. In the Gel Electrophoresis it looks like in lane 2 and 3 there are some non specific amplification, do you know if this is the case or if there is another reason for that result?
Thank you for your response! As for the question you had, those lanes are just filled with the DNA that we isolated from a plaque streak containing both morphologies of plaque. We wanted to continue with experiments involving characterization of the DNA (breaking it down with enzyme restriction digest, and PCR), but ran our of time and could only do a ‘quality control’ experiment to confirm we had DNA present from the plaques. Other than the presence of the DNA being illuminated by SyberSafe, I’m unsure the reason for the amplification. Not sure how much it affects it, but the third lane has more DNA than the second, and could be brighter due to this. Thanks for the question!
Why do you believe your phage exhibited both lytic and lysogenic lifecycles? Is it possible the environment these plates were stored in was conducive enough to have the lysogenic phages enter the lytic lifecycle some of the time (because you mentioned the lysogenic phages could enter the lytic lifecycle under certain environmental conditions)? Or could it be something else?
Mainly, due to the two different morphologies present in our plaque assays/streaks, we believed that our one phage exhibited both lifecycles – or, we thought there was two phages present. You make a great point though that I didn’t consider! It is definitely possible that the environment in which our phage was stored caused that shift, and is really something interesting to study/ponder. If not, I would assume that we failed to isolate the other phage in EM imaging and there were in fact actually two phages. Thank you for your questions!
Where were the samples in lanes 2 and 3 of figure 3 from? It says they are controls, so what was the experimental factor?
Those two lanes are isolated DNA from a phage streak. We wanted to conduct PCR and other characterization tests but due to timing, were unable to run these experiments. The lanes simply represent the presence of our phage’s DNA and confirm to us that we didn’t have some odd contamination (even though the plaques show this as well). Thank you for questions!
Hello Brent!
I really enjoyed your poster board presentation! I showed you had a lot of knowledge of both your research as well as background into the field. I did have one question regarding your figures and results section. In the Gel Electrophoresis it looks like in lane 2 and 3 there are some non specific amplification, do you know if this is the case or if there is another reason for that result?
Look forward to hearing form you soon!
Cristian
Cristian,
Thank you for your response! As for the question you had, those lanes are just filled with the DNA that we isolated from a plaque streak containing both morphologies of plaque. We wanted to continue with experiments involving characterization of the DNA (breaking it down with enzyme restriction digest, and PCR), but ran our of time and could only do a ‘quality control’ experiment to confirm we had DNA present from the plaques. Other than the presence of the DNA being illuminated by SyberSafe, I’m unsure the reason for the amplification. Not sure how much it affects it, but the third lane has more DNA than the second, and could be brighter due to this. Thanks for the question!