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This abstract was last modified on May 2, 2018 at 5:30 p.m..
With the hope of discovering new bacteriophages and expanding our understanding of phage diversity, we searched for phages that could grow on Microbacterium foliorum NRRL B-24224 SEA in the Phage Discovery Lab course, BIOL 105L, at Gonzaga University during the academic year 2017-2018. We purified and characterized 205 phage isolates from enrichment cultures of soil, grass, acorn, moss, bark and water samples. The phages were propagated at room temperature (22-25 deg C) during isolation, purification, and amplification. At this temperature, lawns and plaques could be observed after 24 hours, but plaques continued to grow in size over the next 3-4 days. Compared to previous students' experiences with M. smegmatis phages, we found that a higher percentage of isolates were difficult to work with. A number of us found it difficult to prepare a lysate with a titer greater than 10<sup>8</sup> pfu/ml, a number of phages appeared distorted in TEM images, and a large percentage of lysates produced DNA that appeared to be fragmented or degraded. We report here on our experiences with these experiments, and our efforts to troubleshoot working with M. foliorum phages.