Allelic diversity of 44 microsatellite marker loci originated from the coding regions of specific genes or the non-coding regions of barley genome was analyzed for 19 barley genotypes. Multi-allelic variation was observed at the most of marker loci except for HVM13, HVM15, HVM22, and HVM64. The number of different alleles ranged from 2 to 12 with a mean of 4.0 alleles per micro-satellite. Twenty-one alleles derived from 10 marker loci are specific for certain genotypes. The level of polymorphism (Polymorphic Information Content, PIC) based on the band pattern frequencies among genotypes was relatively high at the several loci such as HVM3, HVM5, HVM14, HVM36, HVM62 and HVM67. In the cluster analysis using genetic similarity matrix calculated from microsatellite-derived DNA profiles, two major groups were classified and the spike-row type was a major factor for clustering. Correlation between genetic similarity matrices based on microsatellite markers and pedigree data was highly significant (r=0.57** ), but these two parameters were moderately associated each other. On the other hand, RAPD-based genetic similarity matrix was more highly associated with microsatellite-based genetic similarity (r=0.63** ) than coefficient of parentage.