The taxonomic situation of Anguis fragilis species is still unclear in Turkey. In order to clarify this situation, we used the DNA sequences of 16S rRNA and cytochrome b genes to analyze the phylogenetic relationship among A. fragilis populations. A total of 13 haplotypes in 16S rRNA dataset and 20 haplotypes in cytochrome b dataset were detected. Kimura 2-parameter genetic distance was found to be 0.012 for 16S rRNA and 0.026 for the cytochrome b dataset. Neighbor joining (NJ) trees were constructed to analyze phylogenetic relationship among specimens and were supported with median joining networks. Results indicate a clear genetic structuring in A. fragilis populations sampled from north of 408 north latitude of Turkey. Both mitochondrial gene sequences successfully detected the intraspecific variation among specimens of different populations. Genetic structuring, correlated with geographic distance, was found to be significant at the specimens sampled from edge populations of peripherally isolated climatic conditions.