In order to localize Mlp (a powdery mildew resistance gene) on barley, 92 F3 families of D7B×5A Pallas cross were evaluated for their resistance to powdery mildew. Semi- automated AFLP and SSR techniques, with ABI prism 377 DNA sequencer were applied to genotype these families. Ninety nine AFLP and 63 SSR primer combinations were employed to recognize the polymorphism between the parents through bulked segregant analysis (BSA). AFLP and SSR primer combinations showed 1 and 3 percent polymorphism between parents, respectively. One AFLP marker, E38- M50 (124), and one SSR marker, Bmac 0316, were found to be associated with Mlp gene on the long arm of chromosome 6. The segregation ratio 3:1 and 1:2:1 for AFLP and SSR markers respectively and the ratio 3:1 for Mlp gene on F3 population verified the co- segregation between Mlp and molecular markers.