The date palm (Phoenix dactylifera) is a dioecious plant, and its sexual characteristics remain indiscernible until it reaches the reproductive stage, typically between 4 to 5 years of age. The cultivar C. madjoole is widely grown in Palestine and neighboring regions. Early identi cation of male and female seedlings could signi cantly bene t date palm cultivation and breeding efforts. This study aimed to con rm the effectiveness of microsatellite markers in distinguishing between male and female date palms within the cv. Madjoole variety.Out of eight microsatellite primers used, four markers were able to determine the sex of the date palm tree. mPdCIR010 produced a male speci c band (325pb) while mPdCIR093 produced a female speci c band (335 pb); on the other side, mPdCIR015 and mPdCIR078 produced unique female bands (160 and 128 pb), respectively. A total of 15 loci were identi ed, averaging 2.1 loci per primer.SSR markers were able to provide a reliable technique for sex determination in date palm (cv. Madjoole) at an early stage. The utilization of SSR markers will not only advance but also facilitate the establishment of commercial date palm cultivation through seed propagation in Palestine and the surrounding region. Consequently, the available date palm genetic resources could be expanded, leading to enhanced national breeding programs. The ability to use mPdCIR093 as a female speci c marker is approved for cv. Madjoole but needs to be validated with other date palm cultivars.