The objective of this study was to find QTL for milk production traits in Czech Fleckvieh cattle on chromosomes 6, 7, 11, 14, and 23 where QTL were previously identified in other dairy cattle populations. Sixteen grandsire families were genotyped for 38 microsatellite markers on the selected chromosomes. A QTL mapping model based on variance component analysis was implemented via restricted maximum likelihood (REML) to estimate QTL positions and their effects. A significant QTL affecting fat percentage was found at the beginning of chromosome 14 (0 cM), near marker ILSTS039. Suggestive QTL associated with milk production traits appeared on other studied chromosomes (BTA6, BTA7, BTA11, and BTA23). This first QTL search on five chromosomes in Czech Fleckvieh population showed several suggestive QTL that can be promising for further studies and contribute to better understanding of genetics of milk production in the Czech Fleckvieh cattle.