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Mango is one of the most important fruit crop of world. Among the Major mango producing countries India is at the top followed by China, Thailand, Mexico and Pakistan. It is the 2nd most important fruit crop in Pakistan. Researchers are working on mango breeding to develop new cultivars using various biotechnological tools. Mango is woody plant so it is very hard to deal with it during In vitro regeneration. Present research was designed to optimize the protocol for in vitro regeneration of mango. Leaves, shoot tips and nucellus of Anwar rataul variety were used as an explant. Explants were collected from the fruit plant nursery at UAF. Murashige and Skoog (MS) media supplemented with different levels of PGR's (IAA, NAA and BAP) was used. Inoculation of explants in test tubes was done in laminar air flow hood chamber and cultures were placed in growth room under controlled light and temperature. This study followed Completely Randomized Design (CRD) with thirteen treatments and three replications. Data was collected for the different parameters of callus induction, rooting and shooting and was analyzed by using standard statistical techniques.