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Real‐time PCR identification of lake whitefish Coregonus clupeaformis in the L aurentian G reat L akes
Author(s) -
Overdyk L. M.,
Braid H. E.,
Naaum A. M.,
Crawford S. S.,
Hanner R. H.
Publication year - 2016
Publication title -
journal of fish biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.672
H-Index - 115
eISSN - 1095-8649
pISSN - 0022-1112
DOI - 10.1111/jfb.12922
Subject(s) - coregonus clupeaformis , amplicon , biology , standard curve , real time polymerase chain reaction , fish <actinopterygii> , microbiology and biotechnology , mitochondrial dna , polymerase chain reaction , fishery , gene , genetics , biochemistry
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 .