Octopus cyanea is an important global fishery commodity that inhabits a wide range of intertidal reefs in the Indo-West Pacific region. It plays a significant role in fisheries and is classified as an economically important species due to its high nutrient content. Understanding species diversity is essential for managing octopus's resources, and effective fisheries management planning strategies are needed, particularly for octopus fisheries. In this research, the standard for species identification used as part of the DNA barcoding framework is the cytochrome c oxidase subunit I (COI) gene. The study aimed to determine octopus species based on phylogenetic analysis of COI mtDNA in the Alas Strait. Octopus sampling was conducted in July 2023 using a 10m long and 3m high octopus fishing pole, known as an octopus pocong. Samples were collected from six locations in the Alas Strait: Pringgabaya, Labuhan Haji, Tanjung Luar, Poto Tano, Labuhan Lalar, and Benete. Tentacle samples were taken by cutting approximately 5cm of the tentacles with a sterile knife, then placed in 96% ethanol and labeled. This research identified two species of octopus: Octopus laqueus and Octopus cyanea. Among the six collection locations, Octopus cyanea was the dominant species. Barcoding results using BLAST with primers LCO1490/HCO2198 demonstrated their suitability for octopus's identification in this study. Overall, this research highlights the feasibility of using COI sequences for species identification, providing an initial dataset for future octopus DNA barcoding, especially in the waters of the Alas Strait.