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Recent decades have seen the fast growth of cephalopod fisheries but their management is compromised by the critical gaps in our knowledge of cephalopod life histories. Molecular markers are invaluable tools for studying the evolutionary significance and management implications of variation in mating systems. We have developed seven polymorphic microsatellite loci for mating system analysis in the southern calamari Sepioteuthis australis Quoy & Gaimard 1833 using magnetic enrichment and colony hybridization techniques. Observed heterozygosities range from 32% to 100% and will have sufficient power to examine the relative success of alternate mating strategies in S. australis.