The invention aims to provide a kit for detecting mutation of mitochondria T12201C related with epicophosis, and the kit consists of a DNA extraction mixed liquor, a PCR mixed liquor for amplifying T12201C segments, a pair of outer primers designed according to T12201C, a pair of inner primers designed according to T12201C, restriction enzyme, a positive control unit, a negative control unit and a kit body. By using a proteinase K digestion cracking method to extract the genomic DNA from a few samples, the kit disclosed by the invention not only solves the problem that the DNA is extracted from blood of an epicophosis patient traditionally, relieves the pain of the detected patient, but also solves the problem of sample transmission between different areas, not only improves the specificity of enzyme digestion but also ensures the specificity and stability of the results, not only lowers the detection cost but also realizes simplicity and quickness in operation, and can be applied to detection of mutation of mitochondria gene T12201C related with epicophosis.