A blocking primer increases specificity in environmental DNA detection of bull trout (Salvelinus confluentus)

Taylor M. Wilcox; Michael K. Schwartz; Kevin S. McKelvey; Michael K. Young; Winsor H. Lowe

doi:10.1007/s12686-013-0113-4

A blocking primer increases specificity in environmental DNA detection of bull trout (Salvelinus confluentus)

Taylor M. Wilcox
, Michael K. Schwartz
, Kevin S. McKelvey
, Michael K. Young
, Winsor H. Lowe

Division of Biological and Biomedical Sciences

Research output: Contribution to journal › Article › peer-review

27 Scopus citations

Abstract

Environmental DNA (eDNA) is increasingly applied as a highly sensitive way to detect aquatic animals non-invasively. However, distinguishing closely related taxa can be particularly challenging. Previous studies of ancient DNA and genetic diet analysis have used blocking primers to enrich target template in the presence of abundant, non-target DNA. Here we apply a blocking primer to increase the specificity of a TaqMan assay for eDNA detection of rare and endangered bull trout (Salvelinus confluentus) in the presence of the closely related (Salvelinus namaycush). We found that addition of a blocking primer substantially increased assay specificity without compromising sensitivity or quantification ability.

Original language	English
Pages (from-to)	283-284
Number of pages	2
Journal	Conservation Genetics Resources
Volume	6
Issue number	2
DOIs	https://doi.org/10.1007/s12686-013-0113-4
State	Published - Jun 2014

Funding

Acknowledgments The authors thank the McCutheon Lab at the University of Montana for use of their spectrophotometer. TW is supported by a NSF Graduate Research Fellowship (Grant No. DGE-1313190).

Funder number
DGE-1313190

Keywords

Blocking primer
Bull trout
Complex templates
Environmental DNA

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10.1007/s12686-013-0113-4

Cite this

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title = "A blocking primer increases specificity in environmental DNA detection of bull trout (Salvelinus confluentus)",

abstract = "Environmental DNA (eDNA) is increasingly applied as a highly sensitive way to detect aquatic animals non-invasively. However, distinguishing closely related taxa can be particularly challenging. Previous studies of ancient DNA and genetic diet analysis have used blocking primers to enrich target template in the presence of abundant, non-target DNA. Here we apply a blocking primer to increase the specificity of a TaqMan assay for eDNA detection of rare and endangered bull trout (Salvelinus confluentus) in the presence of the closely related (Salvelinus namaycush). We found that addition of a blocking primer substantially increased assay specificity without compromising sensitivity or quantification ability.",

keywords = "Blocking primer, Bull trout, Complex templates, Environmental DNA",

author = "Wilcox, \{Taylor M.\} and Schwartz, \{Michael K.\} and McKelvey, \{Kevin S.\} and Young, \{Michael K.\} and Lowe, \{Winsor H.\}",

note = "Funding Information: Acknowledgments The authors thank the McCutheon Lab at the University of Montana for use of their spectrophotometer. TW is supported by a NSF Graduate Research Fellowship (Grant No. DGE-1313190).",

year = "2014",

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doi = "10.1007/s12686-013-0113-4",

language = "English",

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T1 - A blocking primer increases specificity in environmental DNA detection of bull trout (Salvelinus confluentus)

AU - Wilcox, Taylor M.

AU - Schwartz, Michael K.

AU - McKelvey, Kevin S.

AU - Young, Michael K.

AU - Lowe, Winsor H.

N1 - Funding Information: Acknowledgments The authors thank the McCutheon Lab at the University of Montana for use of their spectrophotometer. TW is supported by a NSF Graduate Research Fellowship (Grant No. DGE-1313190).

PY - 2014/6

Y1 - 2014/6

N2 - Environmental DNA (eDNA) is increasingly applied as a highly sensitive way to detect aquatic animals non-invasively. However, distinguishing closely related taxa can be particularly challenging. Previous studies of ancient DNA and genetic diet analysis have used blocking primers to enrich target template in the presence of abundant, non-target DNA. Here we apply a blocking primer to increase the specificity of a TaqMan assay for eDNA detection of rare and endangered bull trout (Salvelinus confluentus) in the presence of the closely related (Salvelinus namaycush). We found that addition of a blocking primer substantially increased assay specificity without compromising sensitivity or quantification ability.

AB - Environmental DNA (eDNA) is increasingly applied as a highly sensitive way to detect aquatic animals non-invasively. However, distinguishing closely related taxa can be particularly challenging. Previous studies of ancient DNA and genetic diet analysis have used blocking primers to enrich target template in the presence of abundant, non-target DNA. Here we apply a blocking primer to increase the specificity of a TaqMan assay for eDNA detection of rare and endangered bull trout (Salvelinus confluentus) in the presence of the closely related (Salvelinus namaycush). We found that addition of a blocking primer substantially increased assay specificity without compromising sensitivity or quantification ability.

KW - Blocking primer

KW - Bull trout

KW - Complex templates

KW - Environmental DNA

UR - https://www.scopus.com/pages/publications/84900830845

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DO - 10.1007/s12686-013-0113-4

M3 - Article

AN - SCOPUS:84900830845

SN - 1877-7252

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SP - 283

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JO - Conservation Genetics Resources

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IS - 2

ER -

A blocking primer increases specificity in environmental DNA detection of bull trout (Salvelinus confluentus)

Abstract

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Keywords

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