This study on Octopus vulgaris focused on the COIII gene region of mitochondrial DNA. Sequences from 21 samples from the Eastern Cape, and 14 samples from the Western Cape, were compared to determine whether different populations exist along the South African coast. A 380-bp segment of the COIII region of mtDNA was amplified using the polymerase chain reaction with specific designed primers. Phylogenetic inference was made using maximum parsimony (MP), maximum likelihood (ML), and distancebased methods. All sequences conformed to a single haplotype. Lack of variation within and between east and west coast samples precluded further population genetic analysis. The sequence obtained in this study was also compared with other sequences lodged in the Genbank database. Phylogenetically, the South African O. vulgaris is closely related to O. vulgaris from Senegal (0.67% divergence) and the Mediterranean (1.51% divergence). Within the Mediterranean group, O. vulgaris from South Africa displayed less sequence divergence from Senegalese and Mediterranean individuals than O. vulgaris from Venezuela (3.85%) and Taiwan (3.87%). These data do not, therefore, refute the hypothesis of a single O. vulgaris genetic population around the coast.