: The objective of this study was to establish the technology to identify barley cultivars by the DNA markers for the purpose of managing the foundation seeds and for protecting the breeder s right and the brand seeds from pirated cultivars. A total of 24 two−rowed barley cultivars, which consisted of 22 Japanese cultivars and 2 foreign cultivars, were identifi ed individually by cleaved amplifi ed polymorphic sequence CAPS analysis of the Restriction Fragment Length Polymorphisms RFLP using 9 sequence tagged site STS primers and 6 kinds of endonucleases, following the electrophoresis of the DNA fragments in 1.8 agarose gels. In addition, we selected 28 CAPS markers, 1 simple sequence repeats SSR marker and 5 randomly amplified polymorphic DNA RAPD markers that could clearly and reliably detect DNA polymorphisms among 24 cultivars.