An investigation was conducted on Fast Protocol for High Frequency in vitro cloning of Banana (Musa acuminata) cv. Grande Naine at the Biotechnology-cum-Tissue Culture Center, OUAT, Bhubaneswar, during the year 2012. This has helped to determine the best media compositions for shoot multiplication and rooting of cv. Grande Naine, so as to get optimum results with a minimized cost of production. MS medium supplemented with 4.0 mg/1 Benzylaminopurine (BAP) and 2.0 mg/1 Kinetin gave the highest number of shoot/explants (11.33) in 30 days. However, MS medium when supplemented with 6.0 mg/1 BAP produced a maximum number of leaves (19.07) with a maximum height 2.73 cm. Among various concentrations of indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) and naphthaleneacetic acid (NAA) for rooting. Half MS medium supplemented with 1.0 mg/1 IBA was found to be ideal for early rooting and producing more number of roots in 21 days. However, MS basal medium was found to be the best treatment to support the formation of long roots. This protocol can be very useful to the future research worker and as well as entrepreneurs for mass production of banana (Musa acuminata) cv. Grande Naine
