*This abstract was presented at the 2019 Spring Engagement Forum @ Radford University*

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Emily Hansen & Dr. Bob Sheehy

Alternative DNA Collection and Species Determination in Amphibians

In this area of vast salamander diversity, difficulties arise in distinguishing

between species, especially those with morphological overlaps. At this time, the widely

used technique to distinguish among, at least to the naive eye, morphologically similar

species is to collect tail or nail clippings of salamanders and use DNA barcoding of the

COI locus to identify the species. These clippings are especially invasive to salamanders,

as clipping body parts reduces energy storage. To solve this problem, we too have been

using DNA barcoding techniques of the COI locus from Plethodon samples, but are

moving away from the invasive sampling. We are developing the process of using buccal

brushes to gather epidermal calls, which then serve as the alternative DNA source used

instead of clippings. Through firm, full-body brushing and treatment with the Puregene

DNA isolation kit, the DNA received from the brushing provides an appropriate

template for PCR amplification of the COI locus. This method is applicable to field

situations for a variety of amphibian taxa, as the DNA collected preserves well and there

is no need for specialized equipment or handling conditions. With more research, we will

be able to identify a wide variety of Plethodon species in the surrounding area, avoiding

mis-identification. This approach should be easily extended to a wide variety of

amphibian taxa.