As fisheries shift towards ecosystem-based management, the need to reduce impacts on by-catch has been increasingly recognised. In this study the catch composition, discard rate, and post-capture survival of species caught by gillnets in Tasmania, Australia, was investigated. Over half the commercial gillnet catch was discarded, with discard rates of ~20% for target and >80% for non-target species. Capture condition, including initial mortality, was assessed using simple criteria for a range of species and related to soak duration. Delayed mortality was also assessed using tank trials and related to capture condition. By combining initial and delayed mortality rates post-capture survival was estimated. Longer soak durations generally resulted in slight, but significant, declines in capture condition and lower initial survival. Nonetheless, when combined with delayed survival, four of the five most commonly caught species (Cheilodactylus spectabilis, Latridopsis forsteri, Aplodactylus arctidens, Cephaloscyllium laticeps) exhibited high post-capture survival (83–100%) for soak durations within the maximum regulated range. Post-capture survival of the one remaining commonly caught species, Notolabrus tetricus, (typically discarded) declined with increased soak duration from 84% to 62%, suggesting that this species would benefit from a further reduction in maximum soak duration. Initial and delayed survival rates for the species retained for tank trials exhibited a significant linear relationship, which was used to estimate delayed survival rates for the rarer species. This method enabled the estimation of potential post-capture survival rates for a diverse range of species and may have application in other data-limited situations where the relationships between fishing practices and by-catch survival are uncertain. Overall, our results suggest that soak duration regulation has been effective in reducing by-catch mortality for many species, noting that some species have low survival rates regardless of soak duration.