An efficient micro propagation protocol was developed for Dalle Khursani, an important chilli cultivar of Sikkim. Aseptic cotyledon, shoot tip and hypocotyl explants of Dalle Khursani (Capsicum annuum) were cultured on Murashige and Skoog medium containing 16 different combinations of plant growth regulators for in vitro regeneration. Regeneration was observed only in 8 combinations of growth regulators among which medium containing 4 mg/l Thidiazuron (TDZ) showed the best result with an average of 2.95 shoots per explant and explant response of 73.95%. This was followed by MS medium containing 4 mg/l TDZ + 0.5 mg/l Gibberrellic Acid 3 (GA3) + 0.5 mg/l Indole Acetic Acid (IAA) with an average of 1.94 shoots per explant and 66.66% explant response. Among the three explants used, cotyledons showed the best response in terms of number of shoots per explant with an average of 1.76. Regenerated shoots elongated and rooted well on MS medium containing 2 mg/l GA3 + 1 mg/l IAA with an average shoot length of 3.10 cm and 6.35 ± 0.98 roots per shoot with explants response of 85% and 75%, respectively. The regenerated plants were acclimatized on a mixture of normal soil and artificial soil with 78% survival.