Abstract: DNA barcoding is a new and efficient method of species identification. In this study,based on DNA barcoding,mitochondrial cytochrome oxidase I gene(COI)and 16S ribosomal RNA(16S rRNA)gene were used as target genes for species identification of sea cucumber samples. The results showed that most sea cucumbers could be identified by a single target of COI or 16S rRNA gene,but some sea cucumbers samples need to be identified by combining two target genes. Species of 24 commercial sea cucumber products were identified by the established DNA barcoding method. 10 samples were consistent with their label,6 commercial sea cucumbers were incorrectly labeled with a phenomenon of low-cost sea cucumbers marked as high-price sea cucumbers;8 samples could not get clear species information based on the label,then these samples were identified by DNA barcoding. The study indicated that DNA barcoding could be applied to species identification of sea cucumbers from market,which will provide technical support for the supervision of commercial sea cucumbers.