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1 November 2009 An Improved Field Method to Obtain DNA for Individual Identification From Wolf Scat
Linda Y. Rutledge, Joshua J. Holloway, Brent R. Patterson, Bradley N. White
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Abstract

Sampling of feces for genetic studies of wild populations can be problematic because of the low quality and quantity of template DNA obtained. We used cotton swabs in the field to isolate the mucous layer on the surface of fresh wolf (Canis lupus, C. lycaon, and their hybrids) scats followed by immediate preservation, and compared microsatellite genotyping of DNA from these fresh field swabs (FS) to that of previously frozen laboratory swabs (LS). In single polymerase chain reactions (PCRs) of 2 multiplexes, amplification at 8 loci was higher in the FS samples (FS  =  50%, LS  =  15%; P  =  0.02) because proportion, quantity, and quality of large fragment wolf nuclear DNA from these samples was greater (2.5–25%, 6.25–62.5 ng/swab, 35% amplified at 1,000 base pairs [bp]) than from the LS samples (1.9%–10%, 4.7–25 ng/swab, 10% amplified at 1,000 bp). Paired blood and fresh field-swabbed samples had identical genotypes. In 84 multiplex PCRs we found no evidence of allelic dropout associated with low template quality or quantity. We conclude that field swabbing of fresh wolf scat facilitates field storage and reduces the need for multiple amplifications at single microsatellite loci, thereby reducing the genotyping costs for wildlife projects that use noninvasive samples.

Linda Y. Rutledge, Joshua J. Holloway, Brent R. Patterson, and Bradley N. White "An Improved Field Method to Obtain DNA for Individual Identification From Wolf Scat," Journal of Wildlife Management 73(8), 1430-1435, (1 November 2009). https://doi.org/10.2193/2008-492
Published: 1 November 2009
JOURNAL ARTICLE
6 PAGES

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KEYWORDS
allelic dropout
Canis spp
DNA quality
fecal sampling
genotyping
methods
noninvasive sampling
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