Salinity is one of the most severe environmental stresses affecting plant productivity worldwide. Studies of the physiological and molecular mechanisms involved in tolerance to salinity indicate that to prevent the accumulation and toxicity of sodium in leaf cytoplasm, plants have developed at least four mechanisms that function in a cooperative manner, i.e., restriction of Na+ influx, active Na+ extrusion at the root-soil interface, controlled translocation of Na+ to the shoots, and compartmentalization of Na+ into the vacuole. Ten barley genotypes showing contrasting phenotypes, selected from the USDA barley core collection (2750 lines) (accessions with high relative biomass under saline stress that have relatively high flag leaf Na+/K+ content, versus accessions with high relative biomass under saline stress that have low flag leaf Na+/K+ content) were cultivated in pots in hydroponic system under greenhouse conditions. Two salt stress treatments were applied to seedlings (7dsm-1 and 15dsm-1) along with control (0dsm-1). All treatments were made in triplicate. The salinity was maintained for the rest of the growth cycle. Primers to amplify the barley candidate genes (HKT1;5, HKT1;4, NHX1, and V-H+-PPase) and to monitor their expression profile by semi-quantitative RT-PCR, were synthesized. This study will contribute to understanding the mechanism of salt tolerance in barley and provide alternative barley genotypes that are tolerant to salinity.