An Improved Field Method to Obtain DNA for Individual Identification From Wolf Scat |
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Authors: | LINDA Y RUTLEDGE JOSHUA J HOLLOWAY BRENT R PATTERSON BRADLEY N WHITE |
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Institution: | 1. Environmental and Life Sciences graduate program, Trent University, Peterborough, ON K9J 7B8, Canada;2. Wildlife Research and Development Section, Ontario Ministry of Natural Resources, Trent University, Peterborough, ON K9J 7B8, Canada;3. Natural Resources DNA Profiling and Forensic Centre, Department of Biology, Trent University, Peterborough, ON K9J 7B8, Canada |
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Abstract: | 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. |
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Keywords: | allelic dropout Canis spp DNA quality fecal sampling genotyping methods noninvasive sampling scat wolf |
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