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Major milk proteins have considerable variants which comes from substitution and deletions in their amino acid sequences. Variants in genes that code for milk proteins, such as β-lactoglobulin (β-LG) have been established as genetic markers for milk production and milk protein composition in dairy cattle. The effect of β-LG variant on milk production traits, such as milk yield, fat yield, protein yield, fat percentage and protein percentage, was estimated for 482 Holstein cows in the first lactation. The β-LG variants were determined by PCR-RFLP technique at the DNA level. Single trait linear model was used for the statistical analysis of the data. Results of this study indicated that β-LG variants affected significantly protein yield (p<0.05) and fat percentage (p<0.05). Animals with the AA variant produced 31kg of milk protein more than animals with the BB variant. On the contrary, cows with the BB variant had fat percentage higher by 0.35 and 0.32% compared with cows with the AA and AB variants, respectively. No associations between the β-LG variants and milk yield, protein percentage and fat yield were found. Therefore, milk production traits could be improved through β-LG typing by increasing the frequency of A variant for protein yield or the frequency of B variant for fat content in Holstein dairy cattle population.