The applicability of simple PCR-based approaches for sex discrimination in the three European Phalacrocoracidae species was tested, using 93 individuals of known sex and two sets of primers (1237L/1272R and 2550F/2718R) for the amplification of the avian sex-specific chromo-helicase-DNA-binding protein gene. We evaluated the accuracy of each set of primers in providing the correct sex for each individual. The first primer set did not produce reliable results. The second provided a band pattern for each sex, easily distinguishable with agarose gel electrophoresis, which correctly identified all the individuals, even in samples of low DNA yield. The amplification products were sequenced and aligned revealing important nucleotide diversity among Phalacrocoracidae species. Compared with morphometric discriminant analysis and DNA-fingerprinting techniques previously applied, the PCR-based sexing with the 2550F/2718R primers is more accurate, less invasive and widely applicable to both adults and chicks, using a variety of DNA sources such as blood, tissue, feathers, egg shells and others.
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