A data-set from the mitochondrial 12S rRNA gene subunit of 11 Australasian lycosid species (six New Zealand species and five Australian species) was generated. Three North American lycosid species, one European species and one New Zealand pisaurid (outgroup) were also sequenced. The sequence data for the 16 species were combined with the published sequences of 12 European lycosids, two Asian lycosids and one Asian pisaurid and were analyzed using parsimony and maximum likelihood analyses. The resulting phylogenetic trees reveal that Australasian species largely form clades distinct from Palearctic and Holarctic species providing further evidence against the placement of Australasian species in Northern Hemisphere genera. New Zealand wolf spiders appear to be related to a subset of Australian genera whereas the other Australian lycosid genera are related to Asian/Holarctic faunas. Gene sequences in the 12S region were useful when examining relationships between closely related genera, but were not as informative for deeper generic relationships.