THE GENETIC diversity of some Egyptian barley cultivars was assessed using twelve primers from the inter-retrotransposons amplified polymorphism (IRAP) marker system. In addition, moisture content, crude, lipids, ash, and crude fiber contents were measured in the studied cultivars. The obtained results showed that the average protein content was 11.19%, and the highest one was recorded for Giza135 cultivar with an average value of 12.3% to a lesser extent in Giza 129 (8.7%). Sixty-three IRAP bands, scores of 33.3% and 66.7% of monomorphic and polymorphic bands were recorded, respectively. The polymorphism heterozygosity index values IRAP marker traits were ranged as H (0.04-–0.47), PIC (0.04–0.36), E (7.50–11.75) HAV (0.00–0.01), MI (0.04–0.07), D (0.04–0.61), and R (0.5.–6.86). The proportion of appearing polymorphism was 63%. This multiplicity was used to measure the difference between the studied cultivars; however, there were no unique bands in all cultivars for each IRAP primer. There was a similarity between the PCA, heat map, and dendrogram distributions of Giza123, Giza124, Giza125, and Giza126. The Bioplot analysis divided the cultivars into four groups. The data revealed a significant difference, among the surveyed varieties of barley cultivars in Egypt, at the molecular level.