During the last decades the ground-breeding Montagu’s harrier (Circus pygargus, Linnaeus, 1758) has changed
its breeding habitats in Europe to agricultural areas in which many local populations would be close to extinction without
a special nest protection regime. Although Montagu’s harrier is a well-studied species in terms of ecology and breeding
biology, its genetic structure and population genetics are almost unknown. As there is a lack of good genetic markers we
developed a set of 19 microsatellite markers comprising 16 new STR markers which were identified by next-generation
sequencing (NGS) using 454 shot-gun pyrosequencing of genomic DNA. The STR markers were arranged into three multiplex
PCR sets for high throughput genotyping and characterised. The marker set provides a powerful tool for kinship
analysis. The combined non-exclusion probability for parent pairs was 1.13*10-11. Only three loci showed PIC values
< 0.50. In total, 121 known family relationships were compared with genetically calculated ones to test the markers
suitability for parentage analysis. In 97.5% of all cases full-sibships were accurately determined and 97.6% of all mothers
were assigned correctly to their chicks. The present multiplex PCR panels can be used to investigate several hypotheses
concerning breeding behaviour, kinship, exchange rates between populations and phylogeography.