Abstract: The identification of many insect species and especially pupae and larval stages can be very difficult by traditional morphological methods as a consequence of high morphological variability within species and due to the existence of sibling species. Coleopteran is the most diverse order of insects and its identification is more difficult and complex. In this study, the PCR techniques was used to amplify 20 common coleopteran larvaes in Qinghai and 566 bp sequences of Mitochondrial Cytochrome Oxidase I( COⅠ) was obtained in order to identify the larvaes which have similar morphology. Sequences were blasted in the GenBank, genetic distances were calculated, and a NJ phylogenetic tree was constructed with Kimmura2parameter distances mode. The results showed that all the larvaes belonged to 5 families and 9 genera based on the analysis of three kinds of methods. In conclusion that DNA Barcoding is a fast and accurate tool for species identification and mtDNA COⅠsequence data can be used as standard DNA barcoding marker for successful species identification of coleopteran insects.