Introduction: The purpose of this study was to study the root canal morphology of South Asian Indian mandibular premolars using a tooth clearing technique. Methods: Two hundred mandibular premolar teeth were collected from different dental schools and clinics in India. After pulp tissue removal and root canal staining with Indian ink, the specimens were decalcified with 5% nitric acid, dehydrated in ethyl alcohol, and subsequently cleared in methyl salicylate. Results: Of the 200 mandibular premolars, 100 were first premolars and 100 were second premolars. Of the first premolars, 94% had a single root, whereas 6% were 2 rooted. Seventy-six percent had a single canal, 22% had 2 canals, and 2% had 3 canals. Eighty-two percent had a single apical foramen, 16% had 2 foramens, and 2% teeth had 3 apical foramens. Eighty percent of teeth had type I, 6% had type II, 10% had type IV, 2% had type V, and. 2% teeth had type IX root canal anatomy. Of the 100 second premolars, 92% had a single root, whereas 8% teeth were 2 rooted and fused. Fifty-eight percent of teeth had a single canal, and 42% had two canals. Eighty-eight percent had a single apical foramen, and 12% had 2 foramens. Sixty-six percent had type I, 30% had type II, and 4% had type V root canal anatomy. Conclusions: A high prevalence of 2 canals was noted in the first and second premolars. Also, 20% of first premolars and 34% of second premolars had a root canal anatomy other than type I.