Development, validation, and evaluation of an assay for the detection of wood frogs (Rana sylvatica) in environmental DNA

• Main Authors: Spangler, Mark A., Huettmann, Falk, Herriott, Ian C., López, J. Andrés
• Format: Dataset
• Language: English
• Published: Springer 2017
• Subjects: eDNA; wood frog; Rana sylvatica; cytochrome B; Alaska; Arctic; Calvin; Fairbanks; North Pole; Institute of Arctic Biology
• Online Access: http://hdl.handle.net/11122/8005

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). 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. Alaska Herpetological Society University of Alaska Fairbanks Department of Biology and Wildlife (Calvin J. Lensink Graduate Fellowship in Wildlife Biology) University of Alaska Fairbanks Institute of Arctic Biology (IAB Summer Graduate Research Fellowship)