An experiment was conducted for standardization of in vitro propagation technique of Bacopa monnieri (L.), a medicinal herb of India. Healthy leaf segments of the herb were used as explants with basic Murashige and Skoog (MS) medium containing various combinations of different growth regulators for callus, shoot and root initiation. The best callus induction percentage (95.47%) was observed on MS + 0.5 mg/L NAA and 2.0 mg/L BAP (T3). The maximum number of shoots (8), shoot length (9.30 cm) and shoot induction percentage (90.48%) was achieved on MS + 3.0 mg/L BAP and 1.0 mg/L Kn (ST4). The maximum number of roots (8) and root length (7) was observed on MS + 1.5 mg/L IAA (RT5). The rooted micro shoots were successfully hardened and acclimatized in green house and subsequently established in soil with survival rate of 90%.