Objective: The beta-thalassemia (beta-thal) is a diverse group of hemoglobin disorders characterized by a reduced or absent synthesis of the globin chains of hemoglobin (Hb) molecule. Hemoglobin beta gene (HBB) is responsible for synthesis of beta globin chain, and has many mutations, changing from a population to other. beta-thal is an important health problem in the Sanliurfa province placed in Southeastern Anatolia Region of Turkey. We aimed to investigate the frequency of the HBB gene NM_000518.4:c.93-21G>A (IVS1-110G>A or g.252G>A), c.25_26delAA (Cd8 -AA or g.75_76delAA), and c.315+1G>A (IVS2-1G>A or g.496G>A) mutations which were the most common in Turkey by using amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). Material and Methods: Blood samples were collected from 51 patients with beta-thal (25 males and 26 females), and DNA was isolated by using salting out procedure. The normal and mutant alleles of the HBB gene c.93-21G>A, c.25_26delAA, and c.315+1G>A were amplified by using amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) method in two distinct PCR tubes. The PCR products were examined on the agarose gel for presence of DNA marker. Results: The frequency of the c.93-21G>A, c.25_26delAA, and c.315+1G>A mutant alleles in 28 of 51 patients was detected as 26.5%, 3.9%, and 3.9%, respectively. Conclusion: This study was the first research of beta-thal mutations in this province, and showed that the c.93-21G>A mutation was the most frequent in this province among three mutations by ARMS-PCR method, and different from those reported in Central Anatolia, Aegean and Mediterranean regions of Turkey. This research may facilitate the implementations of genetic counseling and prenatal diagnosis in the population of Sanliurfa.