The extant hyena species are the remnants of a formerly diverse group of Hyaenidae, which reached its peak about 5 Ma and contained more than 100 fossil species. Hyenas have been the subject of a number of evolutionary and systematic studies during the last two centuries, due in large part to the dramatical loss of its diversity and geography. The phylogenetic studies incorporating both morphological and molecular analysis have yielded different conclusions regarding the evolutionary relationship between extant and extinct hyenas. In this study, partial ancient DNA sequence of cytochrome b gene in mitochondrial genome was successfully retrieved from Pleistocene hyena bones collected from