Background: Glaucoma is the second leading cause of blindness worldwide, and it is associated with increased intraocular pressure and visual field loss. The most common type of glaucoma, primary open-angle glaucoma (POAG), involves progressive optic nerve damage and the death of ganglion cells in adults. Despite the unknown etiology, genetic predisposition plays a significant role in the development of the disease.Objectives: In order to identify the genetic basis of POAG in Zahedan, Iran, three common mutations of the CYP1B1 gene (G61E, R390H, and R469W) were evaluated in this study.Patients and Methods: Forty patients with POAG were recruited from the ophthalmic divisions of Alzahra hospital, which is associated with Zahedan University of Medical Sciences. The CYP1B1 prevalent mutations of p.G61E, p.R390H, and p.R469W were identified in DNA extracted from the blood samples of patients using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique.Results: We identified no mutations in these patients in the three screened positions.Conclusions: To ensure that these genes play no role in the disease, evaluation of the non-coding regions of both the CYP1B1 and MYOC genes is strongly recommended, since other genes are involved in the pathogenesis of glaucoma.