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Development of a quantitative PCR assay for detecting northwest salamander (ambystoma gracile) in environmental DNA samples

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Abstract

We developed a primer and probe based quantitative PCR assay for use with environmental DNA to detect Northwest salamander (Ambystoma gracile), a species endemic to the temperate Pacific coastal region of North America. The assay targets a region in the mitochondrial DNA (mtDNA) D-loop gene. Tests of the assay were performed in silico (using the NCBI BLAST tool), in vitro (using DNA extracted from A. gracile and related species) and in situ (water samples collected from a lake with a known population of A. gracile). This assay will be useful in efforts to monitor the distribution and occupancy of this species.

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Acknowledgements

Tissue samples were provided by the Burke Museum Herpetology Collection at the University of Washington, Museum of Vertebrate Zoology Tissue Collection at the University of California, Berkeley, Jade Keehn and Susan Barnes (ODFW), Kelly Perry (WDFW), Stephen Nyman (HDRinc), Karen Goldberg (Washington State University), Kurt Jenkins (FRESC) and Jamie Bettaso (USFS). Funding was provided by NPS IAA: P20PG00185. Any use of trade, firm, or product names is for descriptive purposes only and does not imply endorsement by the U. S. Government.

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Authors and Affiliations

  1. U.S. Geological Survey, Western Fisheries Research Center, 6505 NE 65th ST, Seattle, WA, 98115, USA

    Marshal S. Hoy & Carl O. Ostberg

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  1. Marshal S. Hoy
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  2. Carl O. Ostberg
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M.H. and C.O. wrote the main manuscript and both authors reviewed the manuscript.

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Correspondence to Marshal S. Hoy.

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The authors declare no competing interests.

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Hoy, M., Ostberg, C. Development of a quantitative PCR assay for detecting northwest salamander (ambystoma gracile) in environmental DNA samples. Conservation Genet Resour 15, 109–112 (2023). https://doi.org/10.1007/s12686-023-01308-4

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  • DOI: https://doi.org/10.1007/s12686-023-01308-4

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