We determined the amount and temporal stability of genetic differentiation among brook cham sampled from five rivers on Cape Race, Newfoundland, with an electrophoretic analysis of 42 protein coding loci. Fish from four of these rivers were analysed for restriction fragment length polymorphisms in mitochondrial DNA (mtDNA). A single mtDNA clone was observed in all rivers sampled, except one, where 47% offish were from a different and relatively divergent clone (0.31 % sequence divergence). In contrast, Cape Race brook charr show large amounts of genetic differentiation at six enzyme coding loci; Nei's genetic distance ranged between 0,020 and 0.048. This differentiation is relatively stable as no significant differences in allele frequencies were detected between fish sampled from two rivers over two consecutive years. The most divergent population based on protein polymorphism is not that with two mtDNA clonal lineages. In contrast to the commonly held view, mtDNA analyses do not necessarily provide greater resolution of population structure than allozyme analyses.