Effects of time and rainfall on PCR success using DNA extracted from deer fecal pellets |
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Authors: | Todd J Brinkman Michael K Schwartz David K Person Kristine L Pilgrim and Kris J Hundertmark |
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Institution: | (1) 211 Irving I, Institute of Arctic Biology, University of Alaska Fairbanks, Fairbanks, AK 99775, USA;(2) USDA Forest Service, Rocky Mountain Research Station, Missoula, MT, USA;(3) Alaska Department of Fish & Game, Division of Wildlife Conservation, Ketchikan, AK, USA |
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Abstract: | Non-invasive wildlife research using DNA from feces has become increasingly popular. Recent studies have attempted to solve
problems associated with recovering DNA from feces by investigating the influence of factors such as season, diet, collection
method, preservation method, extraction protocol, and time. To our knowledge, studies of this nature have not addressed DNA
degradation over time in wet environments, and have not been performed on fecal pellets of ungulates. Therefore, our objective
was to determine the length of time a fecal pellet from a Sitka black-tailed deer (Odocoileus hemionus sitkensis) could remain in the field in a temperate rainforest environment before the DNA became too degraded for individual identification.
Pellets were extracted from the rectum of recently killed deer and placed in an environment protected from rainfall and in
an environment exposed to rainfall. Pellets from each treatment group were sampled at intervals of 2, 7, 14, 21, and 28 days
after deer harvest. DNA was extracted from sampled pellets and individual samples were genotyped using microsatellite markers.
Amplification failure and errors (dropout and false alleles) were recorded to determine extent of DNA degradation. Eighty
percent of samples in the protected environment and 22% of samples in the exposed environment were successfully genotyped
during the 28-day experiment. With no samples being successfully genotyped in the exposed environment after 7 days, our study
showed that rainfall significantly increases degradation rates of DNA from ungulate pellets. |
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