Abstract
A set of 14 microsatellite loci were used for amplification of faecal DNA from capercaillie (Tetrao urogallus). New internal primers were designed for each locus and were employed to perform a seminested PCR approach combined with a multiplex preamplification method. The aim of this process was to increase the quality of the multilocus genotypes and to reduce the amount of sample required. Values of allelic dropout and false alleles found after three repetitions were 8.1 and 1.4% respectively, and 52.63% of the samples amplified for a minimum of 12 loci. Additionally, capercaillie specific sex primers have been designed. These primers give short products suitable for degraded faecal sample amplification. New primers resulted in 87.5% of samples being successfully sexed, a value seven times higher than using the original P2/P8 CHD sexing method.
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Acknowledgments
This work was funded by Grant CN-05-030 from the “Consejería de Medio Ambiente y Ordenación del Territorio e Infraestructuras del Principado de Asturias”. We thank the “Guardería del Medio Natural del Principado de Asturias” for providing the samples. We also thank Ana Corao and Ana Laviada for technical assistance. We thank Sara de Albornoz for the correction and improvement of our English. We are also indebted to two anonymous referees for valuable suggestions to improve the manuscript.
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Trinidad Pérez and José Fernando Vázquez both authors have contributed equally to this manuscript.
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Pérez, T., Vázquez, J.F., Quirós, F. et al. Improving non-invasive genotyping in capercaillie (Tetrao urogallus): redesigning sexing and microsatellite primers to increase efficiency on faeces samples. Conservation Genet Resour 3, 483–487 (2011). https://doi.org/10.1007/s12686-011-9385-8
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DOI: https://doi.org/10.1007/s12686-011-9385-8