DNA marker information is used to identify and distinguish cattle breeds or individual animals. The purpose of this study was to apply the bovine genotypes kit version 1.1/2.1 to bovine DNA samples (National Institute of Animal Science) taken from Australian/American beef (n=148), Holstein beef (n=170) and Hanwoo cattle (n=177) bred in Jeongeub, Jeonbuk, Korea, so that it could distinguish the Hanwoo breed. The bovine genotype kits consisted of 16 ISAG MS markers, which were used to build a database of genotypes in each group. Genotyping results were analysed using the MS tool kit and Phylip programme to create a phylogenetic tree. The GeneClass 2.0 was used to estimate the breed identification. These analyses found that this kit had 100% capacity to distinguish Hanwoo beef, 95.3% capacity to differentiate Australian/American beef and 90% capacity to identify Korean Holstein steer beef. Hence, it was expected that 16 commercial microsatellite markers were useful to categorize the genetic characteristics of Hanwoo breed and to identify Hanwoo individuals and the origin of beef. In particular, it was expected that these markers would be advantageous in discriminating domestic Holstein beef from Australian/American beef.