Background. Naked barley is a promising food crop. To enhance its production, active breeding is required to create productive varieties. The purpose of this study was to test the STS-marker for the Nud1 gene controlling the hulled phenotype, and use it for the production of naked barley hybrids. Materials and methods. Genotyping of 112 F2 hybrids obtained by crossing the naked black variety ‘Jet’ and the hulled white variety ‘Elf’ was carried out using wF2 and kR1, or tR2 primers in the regular PCR mode to amplify the recessive or dominant alleles of the Nud1 gene, respectively, and also in the multiplex PCR mode, which allows simultaneous amplification of both dominant and recessive alleles of the Nud1 gene. The genotyping data were compared with those on phenotypes of hybrids. Results and discussion. The possibility of using multiplex PCR with a set of primers wF2, kR1, and tR2 for identifying dominant and recessive alleles of the Nud1 gene in hybrid material has been demonstrated. However, while the observed number of hybrids homozygous for the recessive allele nud1 almost completely corresponded to their expected number, the clear predominance of homozygotes for the dominant allele Nud1 and the lack of heterozygotes compared to the expected number of hybrids of these groups indicates erroneous identification of some heterozygotes as dominant homozygotes, which must be taken into account during selection of hulled barleys by genotyping. Conclusions. The STS-marker amplified by primers wF2, kR1, and tR2, can be used to select recessive homozygotes nud1nud1 from hybrid populations, however, additional analysis is required for a more reliable identification of heterozygotes and homozygotes for the dominant allele of the Nud1 gene.