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Characterization of a western North American carnivore community using PCR-RFLP of cytochrome b obtained from fecal samples

Bidlack, Allison L., Reed, Sarah E., Palsbøll, Per J., Getz, Wayne M.
Conservation genetics 2007 v.8 no.6 pp. 1511-1513
DNA primers, carnivores, cytochrome b, feces, genes, laboratory equipment, mitochondrial DNA, polymerase chain reaction, restriction endonucleases, restriction fragment length polymorphism, North America
We developed a simple and reliable method to identify carnivore scats to species using PCR and RFLP of a portion of the mtDNA cytochrome b gene, which works for seven of the most common carnivores in western North America. We identified a short (196 bp) polymorphic region of cytochrome b which would be easily amplifiable even from degraded DNA, developed a primer set, and isolated a set of three restriction enzymes (HpaII, DdeI, HpyCH4V) that would identify the seven target species. In order to test whether this protocol would effectively identify scats obtained in the field we collected 243 carnivore scats from 12 sites in the San Francisco Bay area. Eighty five percent (206) of our samples successfully amplified and were subsequently identified to species using our RFLP protocol. We selected 108 of these samples to sequence; our species identifications based on sequencing were identical to those obtained using our PCR-RFLP method. Our PCR-RFLP method is a simple and efficient means to identify carnivore scats to species, eliminating the need for sequencing, which is costly and requires more laboratory equipment. The technique can also be modified depending on the species present at a particular site. It allows for rapid and noninvasive assessment of multiple carnivore taxa and is particularly useful for surveying populations across many sites.