Beta-thalassemia is one of the most common genetic diseases with autosomal recessive inherited patterns in the world and is one of the most common diseases in Iran that exists in all age and sex groups. Determining gene mutations in this disease can be effective in controlling and treating the disease. The present study determined the frequency of polymorphisms of Hinc, RSaI, RDB, and Xmn in patients with beta-thalassemia minor in Ardabil province. Fifty three beta-thalassemia patients referred to the genetic department of Imam Khomeini Hospital were studied. Blood samples were taken to determine the type of gene mutation. PCR samples were genetically evaluated to determine genetic mutations using RDB-Sequence-RFLP-Haplotype methods. A total of 53 samples were examined, of which 56.6% were male and the rest were female. The most positive cases in the first and second ranks were related to XmnI and AvaII enzymes with 73.5% and 60.3%, respectively. The most common mutation extracted in the studied samples with 14 cases (26.4%) was IVS2.1. Among the most common mutations extracted by the RDB method was related to IVS 1.2 with 26.4%. The results of the present study showed that the distribution of genetic mutations in the studied samples can be different from other places. Also, by performing targeted genetic counseling, it is possible to control and prevent the disease in the future.