10 thoughts on “P52 – Koehler

  1. What was the purpose of using restriction enzymes to cut the DNA? How would this help identify phage cluster identity?

    1. Restriction enzymes cut the DNA at specific, short (usually ~6 base pairs) sequences, creating fragments of various lengths depending upon the subject genome. The number of cuts and lengths of the fragments created gives information about the contents of the test sequence and allows comparison to other phage genomes. This comparison allows identification of phages with similar genomes, providing insight into phage cluster/ subcluster.

    1. If PoplarHRT were a viable candidate for phage therapy, it could help treat antibiotic resistant strain Mycobacterium tuberculosis infections as this bacteria is closely related to M. Smegmatis.

  2. Did you have the option to choose which bacteriophage you tested? If so, why did you choose to isolate poplarHRT? What outcomes did you expect prior to the tests that you ran?

    1. Yes, we had plaques from multiple potential phages from plating our enrichment results, but we chose this phage as it appeared to be lytic (from the clear plaque morphology). I expected to be able to determine a cluster from the restriction digest, but degraded DNA produced inconclusive results.

    2. Yes, we had plaques from multiple different phages in the results from enrichment. We chose PoplarHRT largely because it appeared to be lytic based on the plaque morphology. I expected to potentially identify a cluster that our phage would belong to, but degraded DNA produced inconclusive results.

  3. How would insight into the viral genome sequence help to further your research of poplarHRT for the future?

    1. Insight into the phage genome could provide information into which phages it is similar to using sequence similarity and gene order. This can give an idea of potential host range and help classify the phage.

    2. Comparison to other phage genomes and gene order allows classification and insight into the host range.

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