The Date palm (Phoenix dactylifera L.) is perennial, diploid plant found abundantly in the Middle East region. It plays a significant role in traditional as well socioeconomic growth in the Sultanate of Oman. The main focus of the current work is to identify different methods of DNA extractions for date palm leaf and further to study the genetic diversity among the seven different cultivar varieties, within the Sultanate of Oman, using SSR marker. Seven cultivars were obtained randomly from different places in Oman namely Khasab, Naghal, Barni, Madloki, Khenezi, Nasho and Fard. Genomic DNA was extracted using Edwards's method and Maxi kit method (QIAGEN). Fingerprint is established by PCR amplification of DNA samples with five sets of SSR primers to identify the diversity between individual date palm from seven different locations. This study will help the researchers as a tool and the markers for the similarity and differences analysis as well as mapping of date palm in Sultanate of Oman. The findings of the study indicated that five SSR primers tested were able to produce amplicons for date palms from different locations. Percentage of similarity between seven date palm cultivars from different locations was found to be having 74 % significance.