Abstract: Bacopa monnieri is a vegetatively propagated medicinal plant, which is endangered so it is necessary to develop micropropagation protocols to preserve the germplasm and distribute it to new cultivars during cultivation in new areas. Bacopa is cultivated in marshy area such as rice fields; the plant is of commercial importance. In vitro culture of plants has gained importance during recent years because, besides other application, this technique can be used for the rapid multiplication of medicinal plants. The aim of the present work is to determine the culture conditions for micropropagation of this plant. In order to induce the callus formation from Bacopa monnieri and study the in vitro growth conditions of calli. Study the specific growth factor for root and shoot induction, transplantation and acclimatization of explants and isolation of the DNA from explants. Induction of callus from protoplast of Bacopa monnieri and regeneration of callus. Standardization of micropropagation protocol of Bacopa monnieri. In the case of Bacopa monnieri, the micropropagation technique from explants and protoplasts were established. We also raised plantlets from callus and also subjected the DNA for analysis and done RAPD to find the difference between the normal and in vitro plants, the result indicated that the plants are identical. Standardization of surface sterilization protocol, aseptic culture initiation, establishment and multiplication, suitable media for rooting and a suitable protocol for hardening in order to achieve quality transplant. Optimization of medium for callus induction, maintenance and regeneration.