Argali sheep is an ungulate, which inhabits the north, west, south and central regions of Mongolia. There are two major populations (Altai and Gobi) in Mongolia, but their taxonomic classification as subspecies is often disputed among researchers. Furthermore, there is no recent study about the population genetic structure of argali sheep in Mongolia. In the present study, we have investigated genetic diversity and difference between Altai and Gobi argali populations using mitochondrial control region hyper variable segment (HVS) sequence (598bp) and 3 microsatellite markers. Mitochondrial HVS haplotype analysis showed high haplotype diversity (0.982±0.012) and low nucleotide diversity (0.02589). In microsatellite analysis, total of 9 alleles were found across all loci while mean Ho were 0.59±0.13 for Altai and 0.53±0.1 for Gobi populations, indicating low allelic diversity with moderate heterozygosity. Neighbor-joining tree separated haplotypes into two clusters, Altai and Gobi population, implying distinct genetic difference between the two subspecies. Additionally, Pairwise FST and Kimura-2 parameter showed 0.127 and 0.0413±0.0068, respectively. These genetic distance analyses hinted genetic difference between Altai and Gobi populations are in subspecies level. In summary, mitochondrial HVS and microsatellite analysis demonstrated that Altai and Gobi populations had low genetic diversity but might be genetically distinct from each other in subspecies level, suggesting conservation should be separately managed.