Black sapote (Diospyros digyna Jacq.) is a fruit that is rich in benefits and has high economic value. However, the sex of this plant is difficult to determine early, and data on the plant is still very limited. This qualitative quantitative research was conducted to identify the genetic diversity of black sapote based on RAPD marker to determine the sex of black sapote plants. The sample used consisted of seven black sapote accessions with sample codes including P, M1, M2, M3, F1, F2, and F3. This research consists of several stages, namely DNA extraction and PCR amplification using six RAPD primers (OPA 02, OPA 05, OPA 08, OPA 09, OPC 09, and OPD 15), including electrophoresis to visualize amplified DNA. Data were analyzed using NTSYS 2.02 software and dendogram were obtained using the UPGMA method in the SAHN function. The results of the analysis showed that the polymorphism resulting from the seven black sapote accessions ranged from 50–80%, with PIC (Polymorphic Information Content) values ranging from 0.28–0.50. The closest related accessions are M1 with F2, while accession P with F2 has the highest genetic diversity. The OPA 02 primer with a band measuring 380 bp can be used as a putative marker in the determination of the female sex and hermaphrodite of black sapote plants.