The haplotype and allele frequencies for 17 STR loci of Y-chromosome were obtained for 1151 indigenous Ukrainians from 13 regional populations representing the major territorial subdivisions of Ukraine. There were no significant inter-population differences. The genetic subdivisions within Ukraine was revealed between Polesie, western and eastern forest-steppe populations. The highest microsatellite variability was observed along the edges of Ukrainian area – in the Carpathian region, Bukovina, Sloboda Ukraine; the lowest – in Polesie. The average haplotype diversity values are higher in the steppe and forest-steppe zones, than in Polesie and the Carpathians. Forensic parameters were calculated: total haplotype diversity HD = 0,998855, match probability MP = 0.00114508, the discrimination capacity DC = 0,89400521.