This protocol covers most procedures used by the Nasonia (wasp) geneticist, including virgin collection. Nasonia are easily sexed as pupae, so collecting virgins can easily be performed within a window of several days. The female-specific ovipositor is visible at even the earliest pupal stages as a longitudinal ridge on the ventral posterior abdomen. To prolong generations, yellow pupae can be stored at 4 degrees C-8 degrees C for up to 2 mo and adults for 2-3 wk. In the example presented here, haplodiploid crossing methods are used to make a new Nasonia vitripennis mutant homozygous, using an aunt/nephew mating approach. The protocol can be extended to inbreeding strains. Inbreeding depression generally is not a problem in Nasonia unless the mutation itself is female-sterile. Mother/son mating and sib mating are also reasonable approaches to inbreeding.