Pomegranate is an important economic fruit crop associated with several health benefits which is grown in tropical and subtropical regions of the world. Since this crop has been under cultivation for several centuries, by means of evolution and artificial improvement over a long period, many variations have evolved. Improvement in fruit size, quality, resistance to diseases and pest, high yield are some of important breeding objectives. Usually, the easiest assessment of genetic variation is through morphological and phenotypic measures. However, the morphological traits are often influenced by environmental condition, which can affect the estimation of genetic variation and expression of traits. SRAP is a novel molecular marker technique designed to amplify open reading frames (ORFs), which are the essential components of gene sequences. These markers have several advantages over other, such as ease of use, reasonable throughput rate, high polymorphism, low cost and targeting open reading frames (ORFs). Previous studies have shown that SRAP markers are evenly distributed in the genome, and could generate higher polymorphism than markers such as ISSR, RAPD, and SSR. The present work was carried out by using 114 genotypes of pomegranate which were obtained from United State Department of Agriculture. The DNA from these genotypes was isolated using CTAB method.