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

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...

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Bibliographic Details
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
Description
Summary: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)

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