An eDNA-Based SNP Assay for Ungulate Species and Sex Identification

Many processes in wild populations are difficult to study. Genetic data, often non-invasively collected, may provide a solution to these difficulties and are increasingly used to study behavioral, demographic, ecological, and evolutionary processes. Moreover, the improved sensitivity of genetic methods now allows analyses of trace amounts of DNA left by animals in their environment (e.g., saliva, urine, epithelial cells). Environmental DNA (eDNA) thus offers new opportunities to study a range of historic and contemporary questions. Here, we present a species and sex diagnostic kit for studying browsing in a multispecies temperate ungulate assemblage. Using mitochondrial sequences deposited in Genbank, we developed four single nucleotide polymorphisms (SNPs) for identifying four temperate ungulate species. We also sequenced portions of the Amelogenin gene on the Xand Y-chromosomes and developed six SNPs (three on the X-chromosome and three on the Y-chromosome) for sex determination. We tested the SNP assays on high and low quality/quantity DNA samples.