Project Title: Evaluation of Using PCR-RFLP of Cytochrome b to Distinguish DNA Samples from Carnivore Species in the Southeastern U.S.
Mentor: Todd D. Steury, Forestry and Wildlife Sciences
Project Summary: The goal of our research is to evaluate a relatively simple method for identifying carnivore scats (feces) of eastern North America. The method involves extracting DNA from scat and identifying the species that produced the scat by using special enzymes. The study could lead to a rapid, accurate, and inexpensive method of identifying carnivore populations for use in studies of habitat and population size, with minimal disturbance to the animal being studied.
Abstract: The goal of our research is to evaluate a relatively simple method for identifying carnivore scats (feces) of eastern North America. The method involves extracting DNA from scat, amplifying a portion of the mitochondrial DNA cytochrome b gene using polymerase chain reaction (PCR), and identifying the species that produced the scat using restriction fragment length polymorphisms (RFLPs) and gel electrophoresis. The method was developed and tested for carnivores on the west coast by Bidlack et al. (2007). However, the method has not been tested on carnivores in the eastern U.S. Furthermore, the original study omitted some carnivores that can be relatively common, and thus could complicate species identification. The original study was able to distinguish among DNA from coyote (Canis latrans), red fox (Vulpes vulpes), grey fox (Urocyon cinereoargenteus), bobcat (Lynx rufus), puma (Puma concolor), raccoon (Procyon lotor), and striped skunk (Mephitis mephitis). We intend to evaluate additionally the ability of the method to distinguish DNA from domestic dog (Canis lupus familiaris), domestic cat (Felis sylvestrus), black bear (Ursus americanus), eastern spotted skunk (Spilogale putorius), opossum (Didelphis virginiana), and long tailed weasel (Mustela frenata). Scat samples for all of these species have already been collected as part of other, ongoing field studies. Our plan is to use 1) the Bidlack et al. method to identify collected scats to species via RFLP, 2) sequence amplified DNA using a sequencer (either via the on-campus Genomics and Sequencing Laboratory, or an outside lab), and 3) compare sequence results to those obtained with RFLP. With a rapid, accurate, and inexpensive method of identifying species scat, studies of habitat use, population size, and conservation can be done accurately, efficiently, and with minimal disturbance to the animal being studied. |

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