Genetic discrimination among Solea solea and Microchirus azevia by RFLP analysis of PCR amplified mitochondrial DNA fragments

PCR-RFLP analysis was applied to the differentiation between two closely re lated sole species: Solea solea and Microchirus azevia. Amplification of DN A isolated from fish muscle samples was carried out using four primers flan king two different regions of the mitochondrial genome: a 321 base pairs (b p) fragment from the 12S rRNA gene and a 358 bp fragment from the cytochrom e b gene. Restriction endonuclease analysis based on sequence data of these DNA fragments, revealed the presence of polymorphic sites for several endo nucleases. The restriction profiles obtained by agarose gel electrophoresis when amplicons were cut with those enzymes, allowed the identification of Solea solea and Microchirus azevia species.