Quality characteristics of extra-virgin olive oils depend on several factors. In order to study the effects of genotype and growing location on olive oil quality, olives from cv. Coratina, Nocellara, Ogliarola, and Peranzana, picked in four locations of the Apulia region (Italy), were crushed by a three-phase system to produce mono-cultivar extra virgin olive oils that were analyzed for acidity, peroxide value, spectrophotometric indices, total phenolic content, phenolic profile and antioxidant activity. The experimental data concerning peroxide value, spectrophotometric indices, phenolic content and profile and antioxidant activity showed great variability among the cultivars grown in the same location and also among the oils produced with olives of the same cultivar but grown in different locations. For each cultivar, no significant differences were found among locations in terms of acidity and Delta K whereas peroxide value, K-232, and K-270 differ significantly among locations for both Ogliarola and Peranzana cv. Concerning the phenolic content of Ogliarola cv., no differences were highlighted between the locations whereas the phenolic contents of Peranzana significantly changed as a function of the place of growing. On the basis of these results, the statistical multivariate analysis did not allow the classification into homogeneous groups neither of the oils belonging to the same cultivar nor of those obtained from olives picked in the same location.