To assessment molecular and phenotypic diversity for ten barley lines belong to Hordeum vulgare L., ten SCoT primers were used and 12 morphological traits were estimated in two seasons (2018/2019 and 2019/2020). The SCoT primers succeeded in generating reproducible and reliable amplicons. SCoT technique showed that 66.67 % to 100% of polymorphism. The resolving power (Rp) value varied from 4 to 11.40. In addition, the 10 lines were characterized by 41 unique markers (22 positive and 19 negative). B6 had the highest numbers of positive markers (six). According to phenotypic evaluation, the mean squares for genotypes were highly significant for all studied traits from combined data over two seasons. The heritability values in broad sense (h2b %) ranged from 40.63 (100-grain weight) to 99.22 (Days to heading). The P7 gave desired value in four traits (NT/P, NS/P, NG/S and GY/P g) and the other lines showed desired value in one or two trait, thus all traits which detected in the ten lines might be associated with all unique markers distinguished in this study. The inbred line P6 showed the highest number of unique markers (6 positive), one or some of which may be linked with grain filling period (GFP day) trait that showed in line desirable value. Consequently, these markers may be used as selectable markers for genetic improvement of these traits in barley.