The objective of the present study was to determine the genotype and allele frequencies for osteopontin (OPN), prolactin (PRL) and pituitary-specific transcription factor-1 (PIT-1) that are claimed to be associated with milk production traits in cattle in South Anatolian Red (SAR) and East Anatolian Red (EAR) cattle. In this study, 40 cattle for each of SAR and EAR were used. OPN in intron 4, PRL in exon 3 and PIT-1 in exon 6 were amplified by polymerase chain reaction (PCR) and the resulting PCR products were subjected to digestion using BseNI, RsaI and HinfI enzymes, respectively, to determine the single nucleotide polymorphisms (SNPs). Results showed that frequencies of the OPN allele C, PRL allele B, and PIT-1 allele A which are associated with milk production tarits were found low in SAR and EAR. As a result, SAR and EAR cattle breeds have the lower allel frequencies that influence the milk production traits for three genes than high-yielding European dairy cattle breeds.