Jinten hitam (Nigella sativa) is one of the medicinal plants in the family Ranunculaceae, native to the Mediterranean region and several regions of Asia. The seed of Nigella sativa is widely cultivated, distributed, and used in pharmaceuticals, cosmetics, and food industries. Due to similar seed morphology to its potential adulterant such as Corchorus spp., N. sativa seeds are susceptible to adulteration and substitution in markets. Molecular markers have become one of the most reliable methods for the identification and authentication of medicinal plants. The objective of this study was to select the random amplified polymorphic DNA (RAPD) primer for generating authentication methods of Jinten hitam (N. sativa). Genomic DNA was extracted from samples of N. sativa seed, Corchorus sp. seed, and a mixture of both samples. Forty-two random RAPD primers were used in this study. A total of 227 DNA fragments were produced from 37 amplified RAPD primers, out of which 65% were polymorphic. Primer OPK-4 and OPC-12 generated specific fragments in N. sativa, meanwhile, Primer OPB-1, OPL-5, OPM-3, OPD-5, and OPC-12 generated specific fragments for Corchorus. RAPD molecular marker was able to authentication Jinten hitam (N. sativa) and Corchorus sp. using a selected primer. This research was the first report on RAPD primer screening for Nigella sativa authentication from its potential adulterant (Corchorus spp.).