Abstract Increasing soybean production presents a significant challenge within soybean breeding endeavors. The foundation for achieving success in plant breeding lies in comprehensive knowledge of genetic diversity. Thus, this study aimed to assess the genetic diversity among 30 soybean cultivars by employing microsatellite markers. DNA extraction was conducted using a modified version of the Doyle & Doyle method, followed by amplification of each sample with nine pairs of microsatellite primers. Gel visualization of the amplified samples facilitated scoring, distinguishing between polymorphic and monomorphic primers. The analysis unveiled the detection of 59 alleles, varying from 2 to 15 alleles per locus, with an average of 6.6 alleles per marker. The resulting gene diversity averaged at 0.63, with a mean main allele frequency of 0.44. Moreover, the obtained PIC values ranged from 0.18 to 0.92, with an average value of 0.60. Notably, four microsatellite markers exhibited high informativity (PIC > 0.7). Cluster analysis based on genetic similarity coefficients grouped the 30 soybean cultivars into three main clusters, demonstrating the genetic variability within the utilized soybean genetic material.