We developed and describe a qPCR assay for the detection of wood frogs (Rana sylvatica) using environmental DNA (eDNA) sampling. A single primer set was designed to amplify a 115-bp region of the wood frog cytochrome B gene and assessed for target specificity. There was no evidence of amplification in eleven non-target species. We evaluated the utility of the primer set in qPCR assay by conducting geo-referenced eDNA field surveys in Interior Alaska. Results indicate that the assay consistently detects wood frog DNA in the environment to 1.83x10-3 pg/μL. The assay provides a complement to traditional survey methods and can be readily applied in a wider conservation and management context.
1 liter water samples collected around Fairbanks/North Pole for eDNA analysis. Filters were cut in half and DNA isolated via phenol-chloroform from one half. Other half is archived. qPCR (SYBR Green) conducted for detection of wood frog cytB (Rasy_00).
Spangler M, Huettmann F, Herriott I, and Lopez JA. Development, validation, and evaluation of an assay for the detection of wood frogs (Rana sylvatica) in environmental DNA. Conservation Genetics Resources. doi:10.1007/s12686-017-0881-3
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