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The genetic variability of the Pyrenean frog Rana pyrenaica was assessed using DNA sequences of the cytochrome b, COI and 12S rRNA mitochondrial genes. The results show that, despite inhabiting an area of mountainous terrain with high mountain peaks and deep isolated valleys, R. pyrenaica is genetically very homogeneous. The extremely low level of genetic variability observed, even in the fast evolving cytochrome b and COI genes, suggests that R. pyrenaica may have colonized most of its present range very rapidly after the last Würm glaciation from a refuge area in the Prepyrenees. This is the first attempt to establish the level of genetic variability of this endangered Pyrenean endemism and, as a result, it has major implications for its conservation.