Abstract: Objective:To analyze the gene specificities of mitochondrial cytochrome b (Cytb) of velvet antler and cytochrome C oxidase subunit Ⅰ (COⅠ),and to establish the duplex polymerase chain reaction(PCR)technique for identifying the molecular fingerprint characteristics of velvet antler. Methods:A modified alkaline method was used to extract the genomic DNA from the pilose antler of Cervus Nippon Temminck,Cervus elaphus Linnaeus,Rangifer tarandus and New Zealand deer.Based on Cyt b and COⅠ genes,the specific primers were designed with Premier 5.0 software(Cytb1,2 and CO Ⅰ 1,2,3);PCR amplification was carried out by the single and duplex primers,and the best PCR conditions and highly specific primers were determined. Results:The length of genomic DNA fragment extracted by the modified alkaline method was 23 000bp,and the DNA purity was 1.80±0.02;PCR amplification by the single primer did not identify the authenticity of velvet antler;when Cytb1 and COⅠ1 were applied to PCR, the annealing temperature was 58℃,the fragments of 395 bp and 525 bp were specifically amplified from both Cervus Nippon Temminck and Cervus elaphus Linnaeus antler(Jilin,Anhui),but no fragment appeared for the Rangifer tarandus and New Zealand pilose antler. By using the determined extraction method and the optimal PCR condition,the commercial velvet antlers were carried out and the test results were identical with the reality. Conclusion:The duplex PCR technique can distinguish the authenticity of velvet antler from the molecular level.The method is good in specificity,practicability,and simplicity,and has high application value in the identification of velvet antler.

Key words: velvet antler, cytochrome C oxidase subunit Ⅰ, DNA fingerprint, cytochrome b gene, duplex polymerase chain reaction