Molecular and morphological screening analysis was evaluated to study the similarity levels and marker assisted selection associated with resistance to net blotch. Sequence-Related Amplified Polymorphism (SRAP) as a new marker was used on Egyptian barley for net blotch among 10 Egyptian barley cultivars. Five traits during two consecutive seasons and identify their reaction net blotch. The results exhibited a significant difference among studied cultivars for all traits. The highest mean values for all studied traits detected for, Giza 2000 and Giza 132 which they show resistance to net blotch. Molecular marker analysis using six selected primer combinations gave 56 total bands. Primer combination me5+em5 gave the highest polymorphism (100 %) and the highest polymorphic information content PIC was (0.96). However, primer combination me5+em4 generated specific band associated to net blotch with size ~1090 bp. The dendrogram clustered all studied cultivars into two main clusters each includes the most closed cultivars together according their reactions to net blotch. Results showed that the SRAP marker could be efficiently used to assess genetic variation among Egyptian barley and will be useful for barley germplasm management in terms of biodiversity protection and design of new crosses for disease resistance to net blotch breeding program.