The aim of this research was to assess genetic diversity of Korean-Holstein, Korean Hanwoo, and Ugandan-Holstein dairy cattle. DNA was extracted from either blood or hair of Korean-Holstein (n=74), Korean-Hanwoo (n=75) and Ugandan-Holstein (N=77) using AccuPrep® PCR purification kit. The DNA samples were amplified by multiplex polymerase chain reaction, using GeneTrackTM Hanwoo genotyping kit and assayed using ABI genetic analyser 3130XL. Number of alleles, expected heterozygosity (He), observed heterozygosity (Ho), and the polymorphism information content (PIC) were estimated from 10 microsatellite loci in the three breeds. In addition, F-statistics for each of the 10 microsatellites in the three cattle breeds were estimated using fstat version 2.9.3.2 computer program. GENETIX (v.4.02) was used to perform factorial correspondence analysis (FCA) from the allele frequencies and multi-locus clustering was done using STRUCTURE analyses. A total of 124 alleles were detected. The number of alleles per locus varied from eight (TGLA126) to 22 (TGLA122), with an overall mean of 12.2. Expected heterozygosity ranged from 0.617 (SPS115) to 0.854 (TGLA53) and averaged 0.761. Observed heterozygosity ranged from 0.6 (SPS115) to 0.859 (TGLA53); and averaged 0.761. The mean PIC was 0.723; and means of the F-statistics FIT, FST and FIS were 0.077, 0.076 and 0.001 respectively. Although FCA revealed clear differentiation of Uganda-Holstein, Korean-Holstein, and Hanwoo, clustering assignments showed genetic admixture between Ugandan dairy cattle (Uganda-Holstein) and Hanwoo. In conclusion, the allelic variation present at the 10 loci was sufficient to categorize these cattle into distinct breed groups. Keywords: dairy cattle, genetic differentiation, F-statistics, heterozygosity, microsatellite