This study conducted in the genetic laboratory, department of agricultural botany, in cooperation with laboratory of biotechnology, Faculty of Agriculture, Suez Canal University, Egypt during the period from 2014 to 2015. Fife date palm cultivars in the production period were used in this study as females (Barhiisoidy, Samany, Zaghlool, Nabtet Soltan and Sakey). In addition, three males recognized as superior pollinators (Sakey, Samany, and Zaghlool). These experimental trees are grown at private orchard (Al-raghy) at Ismailia governorate, Egypt. This study aimed to searching for sex-private DNA markers for some date palm cultivars grown in Egypt with the help of DNA marker (nuclear microsatellite or SSRs) to make their election more ease and accuracy, moreover; determine the good pollinators for more uses in the genetic improvement for some quality traits of fruits and increase the yield of palm tree. Eight date palm microsatellite markers were used for gender identification. All eight microsatellite markers analyzed were polymorphic, revealing a total of 205 fragments. The highest number of alleles was featured with Zaghlool cultivar (37 fragments). However, Barhiisoidy cultivar generated the lowest number (17 fragments). From total of the fragments, 65 fragments were scored as private/specific alleles (alleles were unique that genotype alone), among them 64 were as a positive markers and one only as a negative marker. This work is one of the earliest reports for study of the gender genetic determination in date palm cultivars utilizing SSR analyses which gave six positive specific markers for males only. In addition, four positive specific markers for females only. And from another side; the polymorphism level across cultivars was 100 % as revealed by SSR markers.