Evaluation of Fecal DNA Preservation: Techniques and Effects of Sample Age and Diet on Genotyping Success

Michael Panasci; Warren Ballard; Stewart Breck; David Rodriguez; Llewellyn Densmore; David Wester; Robert Baker

doi:10.1002/jwmg.221

Evaluation of Fecal DNA Preservation: Techniques and Effects of Sample Age and Diet on Genotyping Success

Michael Panasci, Warren Ballard, Stewart Breck, David Rodriguez, Llewellyn Densmore, David Wester, Robert Baker

Biological Sciences

Research output: Contribution to journal › Article › peer-review

Abstract

Optimal collection and preservation protocols for fecal DNA genotyping are not firmly established. We evaluated 3 factors that influence microsatellite genotyping success of fecal DNA extracted from coyote (Canis latrans) scats: 1) age of scat, 2) preservative, and 3) diet content. Wequantified genotyping success by comparing rates of allelic dropout, false alleles, and failed amplifications among consensus genotypes. We used a panel of 6 microsatellite loci to genotype 20 scat samples, each of which was subjected to 3 age (1 day, 5 days, and 10 days post-deposition) and 3 preservation (DET buffer, 95% ethanol [EtOH], and lysis buffer) treatments. Both sample age and storage buffer had a significant effect on success and reliability. Ethanol and DET buffer preserved fecal samples with similar efficiency, and both were superior to lysis buffer. Our analysis of DNA degradation rates revealed that samples collected as early as 5 days of age yielded DNA that was highly degraded relative t

Original language	English
Pages (from-to)	1616-1624
Journal	The Wildlife Society
DOIs	https://doi.org/10.1002/jwmg.221
State	Published - Aug 4 2011

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title = "Evaluation of Fecal DNA Preservation: Techniques and Effects of Sample Age and Diet on Genotyping Success",

abstract = "Optimal collection and preservation protocols for fecal DNA genotyping are not firmly established. We evaluated 3 factors that influence microsatellite genotyping success of fecal DNA extracted from coyote (Canis latrans) scats: 1) age of scat, 2) preservative, and 3) diet content. Wequantified genotyping success by comparing rates of allelic dropout, false alleles, and failed amplifications among consensus genotypes. We used a panel of 6 microsatellite loci to genotype 20 scat samples, each of which was subjected to 3 age (1 day, 5 days, and 10 days post-deposition) and 3 preservation (DET buffer, 95% ethanol [EtOH], and lysis buffer) treatments. Both sample age and storage buffer had a significant effect on success and reliability. Ethanol and DET buffer preserved fecal samples with similar efficiency, and both were superior to lysis buffer. Our analysis of DNA degradation rates revealed that samples collected as early as 5 days of age yielded DNA that was highly degraded relative t",

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AU - Panasci, Michael

AU - Ballard, Warren

AU - Breck, Stewart

AU - Rodriguez, David

AU - Densmore, Llewellyn

AU - Wester, David

AU - Baker, Robert

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