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Objective: To establish an accurate, rapid and efficient method to identify the characterization of DNA fingerprint from Trionycis carapax by using molecular biology techniques. Methods: Mitochondrial DNA (mtDNA)was extracted from Trionycis carapax by phenol extraction method and salting-out method respectively. The cytobchrome b (Cytb) partial gene from Trionycis carapax was amplified with PCR by designing specific primers and the PCR products were sequenced and blasted in Genbank. Results: Complete 17.3 kb mtDNA was extracted from Trionycis carapax. The Cyt b gene sequence revealed that the 362 bp segments were identical to that described in GenBank accession number AY687385. Conclusion: Molecular biology techniques can be applied to identify the characterization of DNA fingerprint from Trionycis carapax, with achieving the efforts that were rapid, efficient in addition to high specificity.