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Murphy, M.A.; Waits, L.P.; Kendall, K.C.; Wasser, S.K.; Higbee, J.A.; Bogden, R. |
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Title |
An evaluation of long-term preservation methods for brown bear (Ursus arctos) faecal DNA samples |
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Journal Article |
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Year |
2002 |
Publication |
Conservation Genetics |
Abbreviated Journal |
Conservat. Genet. |
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3 |
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4 |
Pages |
435-440 |
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Abstract |
Relatively few large-scale faecal DNA studieshave been initiated due to difficulties inamplifying low quality and quantity DNAtemplate. To improve brown bear faecal DNA PCRamplification success rates and to determinepost collection sample longevity, fivepreservation methods were evaluated: 90%ethanol, DETs buffer, silica-dried, oven-driedstored at room temperature, and oven-driedstored at -20 °C. Preservationeffectiveness was evaluated for 50 faecalsamples by PCR amplification of a mitochondrialDNA (mtDNA) locus (~146 bp) and a nuclear DNA(nDNA) locus (~200 bp) at time points of oneweek, one month, three months and six months. Preservation method and storage timesignificantly impacted mtDNA and nDNAamplification success rates. For mtDNA, allpreservation methods had >= 75% success atone week, but storage time had a significantimpact on the effectiveness of the silicapreservation method. Ethanol preserved sampleshad the highest success rates for both mtDNA(86.5%) and nDNA (84%). Nuclear DNAamplification success rates ranged from 26-88%, and storage time had a significant impacton all methods but ethanol. Preservationmethod and storage time should be importantconsiderations for researchers planningprojects utilizing faecal DNA. We recommendpreservation of faecal samples in 90% ethanolwhen feasible, although when collecting inremote field conditions or for both DNA andhormone assays a dry collection method may beadvantageous. |
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1572-9737 |
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Equine Behaviour @ team @ Murphy2002 |
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6574 |
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Author |
Taberlet, P.; Waits, L.P.; Luikart, G. |
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Title |
Noninvasive genetic sampling: look before you leap |
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Journal Article |
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Year |
1999 |
Publication |
Trends in Ecology & Evolution |
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Trends Ecol. Evol |
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14 |
Issue |
8 |
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323-327 |
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Hairs; Feces; Feathers; Allelic dropout; Individual identification; Conservation genetics; Behavioural ecology; Pilot study; Microsatellites; Probability of identity |
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Noninvasive sampling allows genetic studies of free-ranging animals without the need to capture or even observe them, and thus allows questions to be addressed that cannot be answered using conventional methods. Initially, this sampling strategy promised to exploit fully the existing DNA-based technology for studies in ethology, conservation biology and population genetics. However, recent work now indicates the need for a more cautious approach, which includes quantifying the genotyping error rate. Despite this, many of the difficulties of noninvasive sampling will probably be overcome with improved methodology. |
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0169-5347 |
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Equine Behaviour @ team @ |
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6573 |
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