Barley is a multipurpose diploid (2n = 14) and the most ancient cereal crop. Studying genetic variability and the magnitude of variation in barley is essential for improving the yields and enhancing production. The experiment was conducted on forty-nine six-row recombinant inbred line food barley genotypes in seven by-seven simple lattice design at Woreilu Farmer Training Center in the 2021/22 cropping season to estimate the extent of genetic variability, heritability, and genetic advance among the genotypes. Analysis of variance (ANOVA) revealed that there was a significant difference (p < 0.001) among forty-nine genotypes for all the traits studied. A high PCV and GCV were recorded for grain yield, fertile tiller per plant, and biomass yield. High GCV with high heritability and genetic advance as a percent of the mean recorded on grain yield, fertile tillers per plant, and biomass yield. Therefore considering these traits for direct selection in barely breeding is important.