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  운영자 2007-05-21 11:34:37 | Hit : 14583 | Vote : 1233
Subject   [자료] Experiments in DNA Extraction and PCR Amplification from Bighorn Sheep Feces: the Importance of DNA Extraction Method
Journal of Heredity 2004 95(6):503-509; doi:10.1093/jhered/esh068

Experiments in DNA Extraction and PCR Amplification from Bighorn Sheep Feces: the Importance of DNA Extraction Method

J. D. Wehausen, R. R. Ramey, II, and C. W. Epps

From the University of California, White Mountain Research Station, 3000 East Line St., Bishop, CA 93514 (Wehausen and Ramey); the Denver Museum of Nature and Science, Denver, CO 80205 (Ramey); and the Department of Environmental Science, Policy, and Management, University of California, Berkeley, CA 94720 (Epps)

Address correspondence to J. D. Wehausen at the address above, or e-mail: johnw@wmrs.edu.

Reliability of genotyping is an issue for studies using non-invasive sources of DNA. We emphasize the importance of refining DNA extraction methods to maximize reliability and efficiency of genotyping for such DNA sources. We present a simple and general method to quantitatively compare genotyping reliability of various DNA extraction techniques and sample materials used. For bighorn sheep (Ovis canadensis) fecal samples we compare different fecal pellet materials, different amounts of fecal pellet material, and the effects of eliminating two DNA extraction steps for four microsatellite loci and four samples heterozygous at each locus. We evaluated 192 PCR outcomes for each treatment using indices of PCR success and peak height (signal strength) developed from analysis output of sequencer chromatograms. Outermost pellet material produced PCR results almost equivalent to DNA extracted from blood. Where any inner pellet material was used for DNA extraction, PCR results were poorer and inconsistent among samples. PCR success was not sensitive to amount of pellet material used until it was decreased to 15 mg from 60 mg. Our PCR index provides considerably more information relative to potential genotyping errors than simply comparing genotypes derived from paired fecal and blood or tissue samples. Our DNA extraction method probably has wide applicability to herbivores that produce pelleted feces where samples dry rapidly after deposition.


http://jhered.oxfordjournals.org/cgi/content/abstract/95/6/503
 Prev   [자료] NONINVASIVE GENETIC SAMPLING TOOLS FOR WILDLIFE BIOLOGISTS: A REVIEW OF APPLICATIONS AND RECOMMENDATIONS FOR ACCURATE DATA COLLECTION
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  2007/05/21 
 Next   [자료] 유럽과 남부/중앙아시아에 분포하는 개과동물의 생태 (IUCN/SSC Canid Specialist Group)
운영자
  2007/05/14 


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