title: Optimizing the use of shed feathers for genetic analysis.
abstract: Shed feathers obtained by noninvasive genetic sampling (NGS) are a valuable source of DNA for genetic studies of birds. They can be collected across a large geographical range and facilitate research on species that would otherwise be extremely difficult to study. A limitation of this approach is uncertainty concerning the quality of the extracted DNA. Here we investigate the relationship between feather type, feather condition and DNA quality (amplification success) in order to provide a simple, cost-effective method for screening samples prior to genetic analysis. We obtained 637 shed feathers of the powerful owl (Ninox strenua) from across its range in southeastern Australia. The extracted DNA was amplified using polymerase chain reaction for a range of markers including mitochondrial DNA, ND3 and nuclear DNA, a simple sequence repeat (Nst02) and a portion of the CHD-1 gene (P2/P8). We found that feather condition significantly influenced the amplification success of all three loci, with feathers characterized as 'good' having greater success. Feather type was found to be of lower importance, with good quality feathers of all types consistently producing high success for all three loci. We also found that the successful amplification of multilocus genotypes was dependant on the condition of the starting material and was highly correlated with successful amplification of the sex-linked CHD-1 locus. Samples with low DNA quality have a higher probability of amplification failure and are more likely to produce incorrect genotypes; therefore, identifying samples with high DNA quality can save substantial time and cost associated with the genetic analysis of NGS. As a result, we propose a method for screening shed feathers in order to provide a subset of samples which will have a greater probability of containing high quality DNA suitable for the amplification of multilocus genotypes.
authors.affiliation: School of Life and Environmental Sciences, Deakin University, 221 Burwood Hwy Burwood, Victoria 3125, Australia, Population & Evolutionary Genetics Unit, Sciences Department, Museum Victoria, GPO Box 666, Melbourne, Victoria 3001, Australia, Department of Genetics, University of Melbourne, Victoria 3010, Australia.
authors.forename: Fiona E
authors.initials: FE
authors.lastname: Hogan
authors.forename: Raylene
authors.initials: R
authors.lastname: Cooke
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Burridge
authors.forename: Janette A
authors.initials: JA
authors.lastname: Norman