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¹ Sriwijaya University, Faculty of Agriculture, Program Study of Aquaculture, Jl. Palembang-Prabumulih Km 32, 30862, Indralaya, South Sumatra, Indonesia DOI : 10.4194/2459-1831-v1_2_01 Viewed : 16923 - Downloaded : 8727 Asian Redtail Catfish (Hemibagrus nemurus) is highly economic important catfish in South Sumatra. Investigating DNA authentication is of importance for species conservation and breeding selection. Cytochrome C Oxidase subunit I is one of mitochondrial DNA markers used for species barcoding in freshwater, brackishwater and marine fish. This research aims to explore the use of COI gene for species barcoding, constructing phylogenetic tree of Asian Redtail Catfish (ARC). The methods used in the research consisted of DNA extraction, PCR (Polymerase Chain Reaction) amplification and sequencing mtDNA COI gene of ARC obtained from Musi and Penukal Rivers (South Sumatra). A 572 and 596 base pairs of partial coding sequences were obtained representing each river. Phylogenetic analyses indicated that ARC was at the same cluster from Bagridae family, and different cluster from others catfish, Pangasius sp and Oreochomis niloticus. Further study using more species of Bagridae and habitat are needed to investigate the diversity of DNA from South Sumatra water resources. Keywords : Asian redtail catfish, COI gene, DNA authentication, phylogeny, South Sumatra