Oriental giant squirrels are tree squirrels classified under family Ratufinae. In India, there are three species of genus Ratufa, i.e. Ratufa bicolor, Ratufa macroura and Ratufa indica. They are also distributed in South and Southeast Asia. However Ratufa indica is endemic to India. The fourth species Ratufa affinis is restricted to Maritime Southeast Asia (East Malaysia, Indonesia, Brunei and Thailand) and probably in Singapore. The species is near threatened .The species R.macroura is endemic to South Asia. Forests of South and Southeast Asia are hotspots of squirrel diversity but at the same time they are at a high risk of extinction because of high deforestation rate and habitat fragmentation. The present molecular study is the first study of the species of Ratufa for their identification. In this study old taxidermy samples were used for amplification of 12SrRNA and Cytochrome b genes. Maximum likelihood and neighbor-joining methods were used to delineate the species by using MEGA 6 and also for molecular analysis for variable, conserved, parsimony and singleton sites. Similarities between species through BLAST indicated 92.21-89.57% between R.macroura vs. R. bicolour; 93.22-90% Ratufa macroura vs. Ratufa indica; 96-92% Ratufa indica vs. Ratufa bicolour, 93.88% between Ratufa affinis vs. Ratufa indica, 93.5% R. affinis vs. R. bicolor, 90.5%. R. affinis vs. R. macroura. Ratufa bicolor is noted to be genetically closer to R. indica as inferred by using both markers. The BLAST result indicated that the obtained sequences matched 99-100% with their respective species. It was also noted that R. bicolor of Jalpaiguri, West Bengal are genetically closer to that of Bhutan. The study also revealed the evolution of R. indica and R. macroura from a single population.