Restriction fragment analysis was used to measure mitochondrial DNA(mtDNA) variation within and among five species of the genus Rana in the South Korea. Purified mtDNAs were digested with 11 restriction enzymes. Six different mtDNA size classes, ranged 19.0 to 28Kb, were observed. The mtDNAs of R. amurensis and R. rugosa shown on intraspecific variations in size could attributed an addition or deletion of about 0.5Kb short fragment. R. rugosa had the largest mtDNA size, in which have been reported in vertebrates. Most restriction fragments were homogeneous within the populations of conspecies; however, intrapopulation (interindividual) variations were discovered in a number of enzyme digests of R. nigromaculata, R. plancyi and R. dybowskii mtDNA. The comparisons of restriction fragments permitted the estimation of the levels of fragment homology (F value) and nucleotide sequence divergence (p value). Mean p value within five species was 0.009 (range of 0-0.022) and 0.192(0.092-0.219) among these species. Presumed divergent times, based million years ago. Especially, the degree of mtDNA sequence divergence among five species was higher than that of any other taxa. The patterns of mtDNA sequence divergence for genetic relationship were very similar to those of the nuclear sequence divergence previously estimated from allozyme.