Penguins (Spheniscidae) represent a monophyletic group comprised of 17 species (Stonehouse 1975; Williams 1995). Found exclusively in the Southern Hemisphere, these flightless diving birds occupy a wide range of habitats from Antarctica to the Galápagos Islands (Williams 1995). Currently, ten species are listed on IUCN’s Red List (BirdLife International 2000) as either ‘endangered’ or ‘vulnerable’. Genetic markers would be useful for resolving issues relevant to the conservation of this group. The entire Adélie penguin mitochondrial (mt) DNA control region has been reported (Ritchie and Lambert 2000) but to date, no general spheniscid PCR-primers that target this useful hypervariable sequence exist. Here we report four novel polymerase chain reaction (PCR) primers designed for the mtDNA control region that amplify across a broad range of penguin species. In addition, we show cross-amplification of nuclear microsatellite loci isolated from Adélie penguins in other penguin species.