We know little about the availability of potential prey and patterns of prey consumption by Ningaui yvonneae in a natural environment. This information is important to understanding how the species is able to exploit its semiarid environment. Here, we examine the diet of N. yvonneae inhabiting a semiarid mallee community in South Australia using a combination of faecal analysis and observations of nocturnally foraging animals. Prey consumption was compared with the availability of prey types in the environment, and comparisons made between habitat components, across seasons and between sexes. N. yvonneae was found to include a range of prey types in its diet, with 11 invertebrate taxa recorded from direct observation and eight of these detected in the scats. Prey taxa consumed most often by freely foraging ningauis were Araneae, Blattodea and Orthoptera, while those most commonly detected in scats were Hymenoptera and Araneae. In comparison, 22 invertebrate taxa were recorded in pitfall traps, although many of these were recorded infrequently. Taxa most commonly recorded in traps were Hymenoptera, Collembola, Coleoptera and Acariformes. Observational data also indicated that consumption of prey taxa differed significantly between sexes and among seasons. Males generally consumed a more diverse array of prey, with both observational and scat data showing that they were more likely to consume Hymenoptera and Isoptera, while females were more likely to consume Lepidoptera and Hemiptera. Variability between seasons was evident in consumption of invertebrate grubs (recorded only during ‘growth’; February–April), Orthoptera (recorded most often during ‘breeding’, August–October), and Araneae (recorded more often during ‘maturation’, May–July). However percentage occurrence data from the scats lacked this seasonality, possibly because of the smaller array of prey taxa recorded. N. yvonneae captured prey from all available habitat components, with five of the most frequently recorded prey taxa being recorded from all components. Leaf litter and Triodia were the most commonly recorded capture sites (45% and 22% of captures respectively), and males and females used the habitat components in a similar manner. Although N. yvonneae is like many other dasyurids in having a largely generalist insectivorous diet, comparisons of prey consumed with their availability in the environment indicated that individuals did show some selectivity towards certain taxa, particularly Blattodea, Orthoptera, Chilopoda, Lepidoptera and Araneae.