The investigation was carried out aiming to develop a technique for rapid in vitro micropropagation of potato with three different popular cultivars in middle Gujarat. Different surface sterilization treatments were carried out and axenic cultures were established on agar solidified (0.8%) Murashige and Skoog’s (1962) medium containing 2% sucrose and 10 mgl-1 GA3. The axenic culture of different cultivar reported the difference among them and concluded that a higher concentration is good for vegetative growth whereas the lower concentration favors the root growth in the presence of 2.0 mgl-1 calcium pantothenate. Effect of CaCl2 on the growth of in vitro cultures was evaluated and it was found that Ca nutrition is one of the most important factors for growth. The produced in vitro cultures were used for further production of microtuberisation study.