A total of 20 microsatellite DNA markers were used for genetic characterization and determination of phylogenetic relationships of native cattle breeds of Turkey, including the Anatolian Grey (AG), Anatolian Black (AB), South Anatolian Red (SAR), East Anatolian Red (EAR), Southern Anatolian Yellow (SAY), and Zavot (ZAV). DNA samples were isolated from 271 blood samples using an organic method. Amplified polymerase chain reaction products were separated by capillary electrophoresis and genotypes were determined for 20 microsatellites. A total of 269 different alleles were determined. The lowest (7.80) and highest (10.80) mean allele numbers were observed for the ZAV and SAY populations, respectively. TGLA122 was the most polymorphic locus; however, only 7 different alleles were observed for INRA005. A total of 40 different private alleles were determined. The general FIS values were between 0.034 and 0.123. Due to the close location to the domestication center, higher genetic diversities were observed. The observed genetic diversities and the results of the phylogenetic analyses were in agreement with evolutionary history and the geographical origins of Turkish native cattle breeds.