Abstract: In this study, we collected DNA barcodes based on COⅠ sequences from 36 individuals of 10 species in 7 genera of Ophichthyidae. The mean length of the sequences was 655 bp and the average content of T, C, G and A was 27.50%, 28.10%, 26.00% and 18.40%, respectively. The A+T content was higher than 50% in every species. The average Kimura two parameter (K2P) genetic distances of interspecies, intraspecies and genus were 0.52%, 20.07% and 23.96%, respectively. K2P genetic distance values were found increasing with taxonomic level. Interspecific genetic distance was about 38.59 times higher than the differences of intraspecies, which met Hebert's species identification standard (interspecies genetic distance was more than 10 times of intraspecies). As high efficiency of species identification is demonstrated in the present study by DNA barcoding, we conclude that COⅠ sequencing can be used to identify fish species.