In the Sverdlovsk region, dairy cattle of the Holstein breed is widely used, obtained as a result of long-term use of the global gene pool of Holstein servicing bulls. The assessment of the nutritional value of milk from cows-daughters of different servicing bulls of Holstein cattle genealogical lines and its quality from the point of view of raw materials for the dairy industry is relevant and has practical significance. As a result of the conducted research, it was found that the livestock of cattle in the farm belongs to five lines – these are Vis Back Ideal 1013485 (25.1%), Reflection Sovering 198998 (19.6%), Montwick Chieftain 95679 (18.2%), Pabst Governor 882933 (22.7%), and Annas Adema 30587 (14.4%). Several servicing bulls are used in each line. Milk obtained from cows-daughters of servicing bulls of the Annas Adema 30587 line has high nutritional values of milk and its caloric content is 76.15-77.50 kCal, which is higher than average. The milk of cows-daughters of all servicing bulls from all the studied lines, despite the difference in physical-chemical parameters, met the requirements of GOST 31449-2013 “Raw cow’s milk. Technical specifications” of the highest grade. Thus, the farm breeds highly productive Holstein cattle with high milk quality indicators.