Powdery mildew, a common cereal disease caused by the fungus Blumeria graminis, is a major limiting factor of barley production and genetic resistance is the most appropriate protection against it. To aid the breeding of new cultivars and their marketing, resistance genes can be postulated in homogeneous accessions. Although hybrid cultivars (F1) should be homogeneous, they are often not genetically uniform, especially if more than two genotypes are involved in their seed production or due to undesirable self-pollination, out-crossing and mechanical admixtures. To overcome these problems the accepted method of postulating specific resistance genes based on comparing response type arrays (RTAs) of genetically homogeneous cultivars with RTAs of standard genotypes was substituted by analysing the frequency of response types to clusters of pathogen isolates in segregating F2 generations. This method combines a genetic and phytopathological approach for identifying resistance genes. To assess its applicability six hybrid cultivars were screened and from three to seven with a total of 14 resistance genes were found. Two genes were newly located at the Mla locus and their heritability determined. In addition, three unknown dominant genes were detected. This novel, comprehensive and efficient method to identifying resistance genes in hybrid cultivars can also be applied in other cereals and crops.