TY - JOUR
T1 - Evaluation of Fecal DNA Preservation: Techniques and Effects of Sample Age and Diet on Genotyping Success
AU - Panasci, Michael
AU - Ballard, Warren
AU - Breck, Stewart
AU - Rodriguez, David
AU - Densmore, Llewellyn
AU - Wester, David
AU - Baker, Robert
PY - 2011/8/4
Y1 - 2011/8/4
N2 - 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
AB - 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
U2 - 10.1002/jwmg.221
DO - 10.1002/jwmg.221
M3 - Article
SP - 1616
EP - 1624
JO - The Wildlife Society
JF - The Wildlife Society
ER -