The purpose of this study was to determine effects of jugular infusion of either balanced or imbalanced amino acid mixture on milk production and composition in dairy cows. Eight mid-lactation Holstein cows were randomly assigned to 5-d continuous jugular infusions of saline (CTL), essential amino acid (EAA) mixture prepared on the profile of casein (CSN, 160 g/d), EAA mixture excluding leucine (Leu) (−Leu, 163 g/d) or EAA mixture excluding arginine (Arg) (−Arg, 158 g/d) in a duplicated 4 × 4 Latin square design with 4 infusion periods separated by a 7-d interval period. The basal diet was formulated with corn grain, soybean meal, cottonseed meal, corn silage, alfalfa hay and Chinese wildrye grass hay according to NRC (2001) and supplied 1.6 Mcal net energy for lactation (NEL) and 94.4 g metabolizable protein (MP) per kg dry matter (DM) to meet requirements for lactation. The results showed that the dry matter intake (DMI) and normal physiological status were not affected by amino acid mixture infusions. Compared with CTL treatment, the CSN treatment increased milk yield (14.9%, P < 0.001), milk lactose yield (14.5%, P = 0.001), milk fat yield (16.6%, P = 0.01), milk protein yield (18.2%, P < 0.001) and the contents of αS1-casein (αS1-CN, 11.8%, P = 0.007), β-casein (β-CN, 4.2%, P = 0.035) and κ-casein (κ-CN, 8.5%, P = 0.003). However, the −Leu and −Arg treatments had lower milk yield (6.3%, P = 0.058 and 5.7%, P = 0.073, respectively), milk protein yield (8.8%, P = 0.010 and 8.2%, P = 0.011, respectively) and the contents of αS1-CN (7.3%, P = 0.057 and 8.4%, P = 0.026, respectively), β-CN (4.2%, P = 0.033 and 3.8%, P = 0.048, respectively) and κ-CN (5.8%, P = 0.023 and 7.6%, P = 0.003, respectively) than those of the CSN treatment. Milk lactose yield (5.9%, P = 0.076) tended to decrease when Leu was removed from amino acid mixture infusate. In conclusion, the supply of casein profile can increase milk production in dairy cows, but a deficiency of Leu or Arg had negative effects on milk yield and milk protein yield.