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Multiplex PCR amplification of 13 microsatellite loci for <Emphasis Type="Italic">Aquila chrysaetos</Emphasis> in forensic applications
Authors:Marcela Bielikova  Andrej Ficek  Danka Valkova  Jan Turna
Institution:1.Department of Molecular Biology, Faculty of Natural Sciences,Comenius University,Bratislava,Slovakia
Abstract:The golden eagle (Aquila chrysaetos) is an endangered raptor, which is threatened mainly by illegal egg and nestling robbery. Here we describe a fluorescently labeled, multiplex PCR method using 13 microsatellite markers, which provides a powerful tool for the individual identification and parentage testing of the Golden eagle. This test should be applicable to both forensic analysis and population studies. Fifteen polymorphic loci from A. chrysaetos were cross-amplified. Subsequent PCR condition optimization led to the successful co-amplification of 13 different loci in a single PCR reaction. Fifty samples from wild-living individuals and 89 samples from captive-bred individuals were examined. The results indicated that both populations have similar levels of moderate inbreeding, unsurprising in a small population. This probability of excluding a random individual in parentage analysis was 0.9912 for the wild population and 0.9932 in the captive-bred one in the case that both the individual and its mother were examined together. The probability of identity was estimated to be 3 × 10−8 for the wild and 4 × 10−8 for the captive-bred populations. Given the size of the Slovak golden eagle population, this test should therefore be sufficient to reliably identify individual raptors and assess parentage in both conservation studies and forensic analysis.
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