Improving non-invasive genotyping in capercaillie (Tetrao urogallus): redesigning sexing and microsatellite primers to increase efficiency on faeces samples

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.