RESUMEN
The tule elk (Cervus elaphus nannodes) experienced a severe bottleneck in the 1800s, resulting in low genetic diversity. There is a need for high-resolution genetic assays that can be used to differentiate individual elk, including close relatives, with high confidence. An efficient assay requires multiple markers both polymorphic and that can be amplified in concert with other markers in multiplex reactions. To develop such markers, we employed 150-bp paired-end whole genome shotgun sequencing on an Illumina HiSeq3000 platform to discover dinucleotide microsatellite markers. After preliminary screening of these markers, we selected and screened 15 candidate loci and 5 existing tetra nucleotide markers in 56 tule elk. We combined these markers in 2 multiplex reactions and report primer concentrations and PCR conditions enabling their efficient amplification.
Asunto(s)
Ciervos/clasificación , Ciervos/genética , Repeticiones de Microsatélite , Polimorfismo Genético , Alelos , Animales , Femenino , Frecuencia de los Genes , Genotipo , Secuenciación de Nucleótidos de Alto Rendimiento , MasculinoRESUMEN
UrsaPlex is a forensic quality 5-dye multiplexed tetranucleotide STR and sex identification panel for individual genetic identification of North American black bears (Ursus americanus). The panel is validated for the identification of black bears involved in human-wildlife conflict events and poaching investigations. This is the first single multiplex panel composed solely of tetranucleotide STRs derived from black bear and bear-specific sex markers. UrsaPlex produces complete genetic profiles from as little as 78 pg of DNA template and has a probability of identity of 2.63 × 10-13. The panel has also been tested for utility in other ursids, and our results indicate with minor modifications, UrsaPlex should prove valuable in identification investigations involving these species as well.