The objective of this study is to distinguish between male and female of Simmondsia chinensis by a mean of the detection of DNA variation using Random Amplified Polymorphic DNA (RAPD) markers in order to identify differences. - Callus tissue was produced from both male and female jojoba leaves using plant tissue culture technique . Genomic DNA of jojoba plant was extracted using CTAB method. The concentration of DNA ranging between (100-150) µg and0.7g weight of male fresh plant, female fresh plant, male callus, and female callus with purity ranging from (1.01-1.19). In this study, eleven different random primers were evaluated for their usefulness in detecting DNA variation between male and female of jojoba. This involved first optimization of RAPD reaction condition including, DNA template ,Taq polymerase ,and primers . The results obtained were as follows: A callus produced from Simmondsia chinensis cultured on MS medium, using hormone combination of (0.5 mg/l BA, 2.5 mg/l 2,4-D ). Six primers showed no amplification products between jojoba male and female. Five primers (A1, A10, C5,D20,E7) showed amplification and represent a mean of distinguishing between male and female, using these primers wre scored by the presence or absence of DNA bands and the differences in their molecular weight. The results clearly demonstrated the feasibility of using RAPD-PCR in distinguishing between jojoba male and female.