title: Real-time PCR identification of lake whitefish Coregonus clupeaformis in the Laurentian Great Lakes.
abstract: The purpose of this study was to develop a real-time PCR assay to specifically identify lake whitefish Coregonus clupeaformis in larval fish assemblages based on a 122 bp amplicon from the mitochondrial genome. The efficiency of the reaction, as calculated from the standard curve, was 90.77% with the standard curve having an r(2) value of 0.998. Specificity of the assay provided single melt peak in a melt-curve analysis and amplification of only the target species. The assay successfully identified target DNA in as low as 0.1% proportion of a DNA mixture. This assay was designed on the portable Smart Cycler II platform and can be used in both field and laboratory settings to successfully identify C. clupeaformis.
authors.affiliation: Department of Integrative Biology, University of Guelph, 50 Stone Road East, Guelph, ON, N1G 2W1, Canada.
authors.forename: L M
authors.initials: LM
authors.lastname: Overdyk
authors.affiliation: Institute for Applied Ecology New Zealand, Auckland University of Technology, Private Bag 92006, Auckland, New Zealand.
authors.forename: H E
authors.initials: HE
authors.lastname: Braid
authors.affiliation: Department of Integrative Biology, University of Guelph, 50 Stone Road East, Guelph, ON, N1G 2W1, Canada.
authors.forename: A M
authors.initials: AM
authors.lastname: Naaum
authors.affiliation: Department of Integrative Biology, University of Guelph, 50 Stone Road East, Guelph, ON, N1G 2W1, Canada.|Chippewas of Nawash, Unceeded First Nation, 135 Lakeshore Blvd, Nevaashiinigmiing, ON, N1G 2W1, Canada.
authors.forename: S S
authors.initials: SS
authors.lastname: Crawford
authors.affiliation: Department of Integrative Biology, University of Guelph, 50 Stone Road East, Guelph, ON, N1G 2W1, Canada.|Biodiversity Institute of Ontario, University of Guelph, 50 Stone Road East, Guelph, ON, N1G 2W1, Canada.
authors.forename: R H
authors.initials: RH
authors.lastname: Hanner
mesh_terms: D000818:Animals; D058893:DNA Barcoding, Taxonomic; D017931:DNA Primers / Q000235:genetics; D004272:DNA, Mitochondrial / Q000235:genetics; D006114:Great Lakes Region; D060106:Lakes; D060888:Real-Time Polymerase Chain Reaction; D012487:Salmonidae / Q000145:classification*; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D004272:DNA, Mitochondrial
keywords: Coregoninae; DNA barcoding; TaqMan; entrainment; species identification