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A system for sex determination from degraded DNA: a useful tool for palaeogenetics and conservation genetics of ursids
Authors:Marie Pagès  Célia Maudet  Eva Bellemain  Pierre Taberlet  Sandrine Hughes  Catherine Hänni
Affiliation:1. Paléogénétique et Evolution Moléculaire, Institut de Génomique Fonctionnelle de Lyon, Université de Lyon, UMR 5242 CNRS, INRA, Université Claude Bernard Lyon 1, Ecole Normale Supérieure de Lyon, 46 Allée d’Italie, 69364, Lyon Cedex 07, France
2. Institut des Sciences de l’Evolution, CNRS UMR 5554, Université Montpellier II, Place Eugène Bataillon, 34095, Montpellier Cedex 05, France
3. Laboratoire d’Ecologie Alpine, CNRS UMR 5553, Université Joseph Fourier, B.P. 53, 38041, Grenoble Cedex 09, France
Abstract:
In this paper, we characterise three sex-specific genes (ZFX/Y, SRY, AMLX/Y) for all eight extant bear species and propose a new, robust and accurate molecular procedure to identify the sex of bears from non-invasive samples and fossil remains. These materials contain tiny amounts of poorly preserved deoxyribonucleic acid (DNA), leaving Polymerase Chain Reaction (PCR) amplification very prone to contamination and difficult to analyse. By taking into account the ancient DNA requirements, the duplex procedures that we developed are efficient not only on DNA extracted from bear faeces but also on ancient DNA extracted from a brown bear fossil 7,500 years old. Defined specifically for ursids, the procedure for faecal samples (co-amplification of ZFX/Y and SRY markers) appears more accurate than other published procedures, as it prevents cross-amplification of potential ingested prey and contamination (19 non-ursid species tested). This system can be applied to threatened bear populations to improve the reliability of sex-ratio and population-size estimates based on non-invasive samples. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorised users.
Keywords:Molecular sex identification  DNA degradation  Non-invasive sampling  Ancient DNA  Ursidae
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