An evaluation of long-term preservation methods for brown bear (Ursus arctos) faecal DNA samples
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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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- An evaluation of long-term preservation methods for brown bear (Ursus arctos) faecal DNA samples
Volume 3, Issue 4 , pp 435-440
- Cover Date
- Print ISSN
- Online ISSN
- Kluwer Academic Publishers
- Additional Links
- DNA extraction
- DNA preservation
- faecal DNA
- noninvasive genetic sampling
- Ursus arctos
- Author Affiliations
- 1. Department of Fish and Wildlife Resources, University of Idaho, Moscow, ID, 83843, USA
- 2. USGS Northern Rocky Mountain Science Center, Glacier Field Station, Glacier National Park, West Glacier, MT, 59936-0128, USA
- 3. Center for Conservation Biology, Department of Zoology, University of Washington, Box 351800, Seattle, WA, 98195, USA
- 4. Amplicon Express, 1610 NE Eastgate Blvd., Pullman, WA, 99163, USA