Abstract: The Karayaka sheep is the most populous sheep breed in the Black Sea region of Turkey and a significant portion of the breed (30-35%) has been raised for centuries in Samsun province. For this reason, we aimed to investigate the genetic diversity of Karayaka sheep raised in Samsun, Turkey. The blood samples used in this study were collected from unrelated animals and were analysed using nine microsatellite markers. A total of 72 alleles were determined with an average of eight alleles per locus. The highest number of alleles (12) was observed for locus BM1314, while the lowest (4) was observed for locus CSSM47. The expected heterozygosity value (He) ranged from 0.654 at locus BM757 to 0.867 at locus BM1314, with an average of 0.757, whereas the observed heterozygosity (Ho) ranged from 0.125 at locus CSSM47 to 0.438 at locus BM757 and HUJ616, with an average of 0.329. The Wright’s fixation index (FIS) values ranged from 0.360 at locus BM757 to 0.833 at locus OARFCB304, with an average of 0.572. The results obtained from this study showed that Karayaka sheep population has a high level of genetic diversity and the studied markers were highly informative.