The essential oil composition and genetic variability of six commercial cultivars of thyme (Thymus vulgaris L.), a Mediterranean medicinal and aromatic plant, were analyzed by GC-MS and randomly amplified polymorphic DNA (RAPD), respectively. All evaluated cultivars belong to the thymol chemotype, with differences in the concentrations of thymol, gamma-terpinene, p-cymene, and other minor components. The comparison of the oil components concentration by multivariate analysis allowed separation of the cultivars into two groups. All of the cultivars exhibited characteristic RAPD patterns that allowed their identification. On the basis of the RAPD patterns, the cultivars could be divided into two clusters, which coincides with results obtained by oil GS-MS analysis, with a correlation coefficient of -0.779.