Data regarding genetic diversity of Arabian oryx(Oryx leucoryx) inWadi Rum area of Jordan is not available so far. Therefore, the current study aimed to evaluate the genetic composition and structure of the Arabian oryx(Oryx leucoryx) herd in Wadi Rum area of Jordan using microsatellite marker analysis.Forty-nine Arabian oryx individuals were captured using a dart gun. Blood samples were obtained via jugular vein puncture into EDTA-blood collection tubes. Then, DNA was extracted and processed using StockMarks® Animal Genotyping System containing 11 microsatellite markers namely, TGLA 227, BM2213, TGLA53, ETH10, SPS115, TGLA 126, TGLA 122, INRA 23, ETH3,ETH225 and BM1824. Then, the samples were analyzed using ABI 310 Genetic Analyzer. Results obtained were used to estimate the level of heterozygosity and allelic diversity and calculation of allelic frequencies of all microsatellites markers, the mean observed and expected heterozygosity, the fixation index, gene flow and Shannon information index using the software GeneAlEx. In the current study the observed Heterozygosity, Expected and Unbiased Expected Heterozygosity were 0.535, 0.431 and 0.435, respectively. These numbers along with the Shannon's Information Index (0.751) indicates high level of within population genetic diversity. High level of genetic variation among Arabianoryx population of the current study ensures genetic diversity in this population which implies that the Arabianoryx population of the current study is almost likely a naturally living wild population surviving on its own.