Sex identification of wolf (Canis lupus) using non-invasive samples
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We have developed new specific primers for sex determination from forensic samples of wolves (Canis lupus), such as hair, saliva, faecal, tooth and urine samples. In order to improve molecular sexing, we performed a multiplex semi-nested polymerase chain reaction (PCR) and several replicated amplifications per sample to avoid errors in low quantity DNA samples, such as allelic dropout and false alleles. The sex of individuals is automatically determined by capillary electrophoresis with a fluorescently labelled internal sex-specific primer from each pair. Our method yielded sex identification on 100% of invasive samples and 93% of forensic samples, being one of the highest success rates obtained from wild animals.
KeywordsCanis lupus Wolf Non-invasive samples Sex-specific primers Sex identification
We are grateful to the personnel from the Central Forest National Reserve (Zapovednik, Russia) for supplying tissue and tooth samples from wolves. We thank “Servei de Protecció de la Fauna, Flora i Animals de Companyia”, “Area d’ Activitats Cinegètiques”, “Guardes de Reserva de Fauna”, “Parc Natural del Cadí-Moixerò” “Forestal Catalana S.A” and “Servei de Diagnòstic Serològic de Leishmania - Servei d’Anàlisi de Fàrmacs, UAB” for making available faecal, urine and hair samples from wolves and dogs. Thanks are due also to Gary Walker for linguistic revision.