Barley is ranked fourth in worldwide cereal production, and is used for a variety of end uses, including animal feed, human consumption and malting. The economic importance of barley has meant that marker- assisted breeding approaches are of considerable interest. Deployment of genetic markers that predict the phenotypic trait of interest with 100 % accuracy (‘perfect markers’) allows efficient tracking of favorable genetic variants through the breeding process, without the need for phenotypic evaluation. Several modern molecular techniques are now being applied together with morphological studies to investigate genetic diversity and relatedness in crops. Various different molecular markers have been used for genetic diversity studies and molecular breeding in barley. These were particularly true for the marker assays such as RAPD (random amplification of polymorphic DNA), AFLP (amplified fragment length polymorphism), restriction fragment length polymorphisms (RFLPs), and STS (sequence tagged site) are the most known molecular markers used for barley breeding.