Triglidae is a family of benthic marine fishes known as gurnards and sea robins, and are distributed throughout all the tropical and temperate seas. Electrophoresis of PCR-RFLP and SDS-proteins were used for differentiation between three gurnards and to detect their genetic relationship. Twenty four fish samples of the grey, the red and the piper gurnards were taken as eight samples from each species. Protein bands varied from 10 to 13 bands in the studied fishes, i.e. 13 bands in the grey gurnard and 10 bands in each of the piper and the red gurnards. The red gurnard recorded the highest similarity to the piper gurnard, while the red gurnard recorded lower similarity to the grey gurnard. Two primers were used for PCR amplification of 360 bp long fragments of the mt DNA cytochrome b (Cyt b) gene and 655 bp of the mt DNA cytochrome oxidase subunit I (COI) gene. The PCR products were digested by two different restriction enzymes DdeI and HincII. Digestion by these enzymes produced highly polymorphic restriction profiles. This enables the differentiation of these fishes and showed that there is a wide intraspecific COI and Cyt b genes. The phylogenetic tree based on the sharing protein bands and PCR-RFLP fragments gave similar results and revealed the relationship between the studied gurnards.