The number of abandoned dogs is increasing with the worsening of the economy and the rising of feed value. It was becoming a serious social problem because of the disease transmission and destruction of natural ecosystems by abandoned dogs been wild animal. In order to solve these problems, companion dogs necessary to secure its own genetic information and to establish the systematic tracking system. Using multiplex-PCR method with 27 microsatellite marker (MS marker) divided 3 set, various alleles occurring to 6 dog breed (Labrador Retriever, German Shepherd, English Springer Spaniel, Belgian Malinois, Jindo Dog, PoongSan Dog) make use of markers to determine allele frequency and heterozygosity. MS marker FH2834 and FH2790 have only two allele and most were found in 13 alleles at FH3381 and FH3399. Average heterozygosity of MS marker is 0.534 and especially, heterozygosity represented the highest value of 0.765 at FH3381. So, it was recognized appropriate allele frequency for individual identification and paternity diagnosis in companion dogs. Using multiplex-PCR method with MS marker, various alleles occurring to dog breed make use of markers to deter mine individual identification and paternity diagnosis, traits associated biomarkers and breed-specific marker for faster, more accurate and ways to reduce the analysis cost. Based on this result, a scientific basis was established to the existing pedigree data by applying genetics additionally. Animal registration system is expected to be conducted nationwide in future. The method expects to very useful this system.