Sorghum (Sorghum bicolor (L.) Moench.) is one of the unique crops existing on its capabilities as a potential source of food, feed and fodder. Genetic factors can be tested by molecular markers which can also shed light on the questions concerning crop quality management. A study was conducted to evaluate the utility of ISSR markers as a tool for identification of genetic diversity among sorghum genotypes. 25 ISSR primers were used to study the genetic diversity among 16 genotypes of Sorghum bicolor (L.) Moench. A total of 288 scorable ISSR markers were generated from 16 genotypes, of which 262 were polymorphic. The polymorphic data generated from these markers were used to estimate the similarity index value and a dendrogram was constructed. The Dendrogram classified the 16 genotypes into 13 distinct clusters. Percent polymorphism observed was 90.97. Based on the results of these studies, ISSR markers appear to be particularly useful for the estimation of genetic similarity among diverse genotypes of sorghum. This book will helps the sorghum breeders to get insight in to the use of marker technology for crop improvement.