Horizontal gene transfer allows for genes that code for certain proteins, like those that allow for bioluminescence, to be added to a phage genome, and then integrate into the bacterial host cell. This would allow for the infected bacteria to light up which makes diagnoses for certain bacteria much easier (Kumar, 2008).
1:10 dilutions were used in this experiment because when calculating the titer of our HTL, we had to use the equation (pfu/µL)(1000µL/1mL)dilution factor, and the 1:10 dilution allows for the dilution factor to be a factor of 10. Additionally, 1:10 dilutions were used earlier in the experiment to ensure that a single phage was isolated from the enrichment of the original soil sample. A different dilution would likely have produced a similar result, as the amount of plaques on the plaque assay plate would scale with the level of dilution that was performed.
For sure! So we first collected a soil sample, which is a common place to find phages, and we enriched this with M. smegmatis bacteria and LB media to allow the bacteria to proliferate and to allow any phages in the soil to infect all these bacteria. We then filtered and collected the liquid obtained from this experiment plated it with M. smeg to determine the presence of a phage. We then collected phages from a plaque on this plate and mixed them with phage buffer, and we diluted this buffer in 2 rounds of serial dilutions to isolate a single phage.
A key limitation in this research was that not every phage in our class could be submitted for sequencing due to the cost, and sequencing is the only way to 100% verify the cluster your phage. So while the PCR result gave us a pretty good indication that our phage is likely part of cluster B1, we can’t be completely sure since we didn’t get to sequence our phage.
curesymposium.org 
How does horizontal gene transfer impact the phage’s usefulness as a diagnostic tool?
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Horizontal gene transfer allows for genes that code for certain proteins, like those that allow for bioluminescence, to be added to a phage genome, and then integrate into the bacterial host cell. This would allow for the infected bacteria to light up which makes diagnoses for certain bacteria much easier (Kumar, 2008).
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Why was a 1:10 dilution used for this experiment, would a different dilution affect the results?
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1:10 dilutions were used in this experiment because when calculating the titer of our HTL, we had to use the equation (pfu/µL)(1000µL/1mL)dilution factor, and the 1:10 dilution allows for the dilution factor to be a factor of 10. Additionally, 1:10 dilutions were used earlier in the experiment to ensure that a single phage was isolated from the enrichment of the original soil sample. A different dilution would likely have produced a similar result, as the amount of plaques on the plaque assay plate would scale with the level of dilution that was performed.
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Can you briefly explain the process how you isolated your phage?
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For sure! So we first collected a soil sample, which is a common place to find phages, and we enriched this with M. smegmatis bacteria and LB media to allow the bacteria to proliferate and to allow any phages in the soil to infect all these bacteria. We then filtered and collected the liquid obtained from this experiment plated it with M. smeg to determine the presence of a phage. We then collected phages from a plaque on this plate and mixed them with phage buffer, and we diluted this buffer in 2 rounds of serial dilutions to isolate a single phage.
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Great presentation! What are some potential limitations to your research?
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A key limitation in this research was that not every phage in our class could be submitted for sequencing due to the cost, and sequencing is the only way to 100% verify the cluster your phage. So while the PCR result gave us a pretty good indication that our phage is likely part of cluster B1, we can’t be completely sure since we didn’t get to sequence our phage.
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