The research was conducted in the laboratory of molecular genetic expertise in 2018 and 2019 within the framework of the state assignment on the topic "Development of genetic-selection and theoretical foundations for the conservation and effective use of the gene pool of cattle in the Ural region with the use of modern biotechnologies". The object of the study is the cattle of Holsteinized black-and-white breed. The main task is to analyze the degree of influence of the somatotropin (GH) gene polymorphism on lactation performance indicators. In the course of the study, DNA from 270 cows was isolated, the polymorphism of the somatotropin gene was determined based on the obtained data, and the relationship between different genotypes and indicators of live weight, milk yield, protein mass fraction and fat mass fraction in milk was established. The results were biometrically processed using IBM SPSS Statistics 23 software program. It was revealed that LL is the main genotype of the GH gene among the representatives of the Holsteinized black-and-white breed (frequency of occurrence 77.0%). The proportion of LV and VV genotypes in the population is 21.9 and 1.1%, respectively. During the observation period, cows carrying the LL genotype showed higher milk yields in the first and third lactations. The difference with their peers was 202-2334 kg (p ≤ 0.05). A tendency towards increased protein content in milk was recorded in animals with the VV genotype. In the first lactation, the difference with LL and LV genotype carriers was 0.03 and 0.06%, respectively. In the third lactation, individuals with the VV genotype surpassed their female counterparts in protein content by 0.07 and 0.12%, respectively.