To cite this paper: Hua, L. Y. and Z. Minghai, 2017. The genetic diversity of tibetan red deer populations determined using mitochondrial and microsatellite markers. We performed genetic analysis using mitochondrial and microsatellite markers. The mtDNA CytB genes from 54 individual Tibetan red deer were sequenced and 731 bp fragments were obtained. We defined 14 haplotypes with a haplotype diversity (H) of 0.897. The nucleotide diversity (π) between populations was 2.781. Microsatellite typing analysis showed that the average allele number, mean expected heterozygosity and mean observed heterozygosity were 4.385, 0.721 and 0.641, respectively. Our findings suggest that these Tibetan red deer populations have relatively high genetic diversity. As indicated by Tajima′s D and Fu and Li′s D values, these three populations had not significantly deviated from a neutral evolutionary path (p > 0.1), nor was any evidence found to suggest a strong equilibrium selection. AMOVA revealed that the inter-gene flow of these three populations was greater than 1 (5.14 > Nm > 1.67), suggesting that abundant gene flow exists between them. No obvious genetic differentiation was observed. Thus, we propose to protect and control the Tibetan red deer in these three regions as a single management unit.