The mussel Mytilus galloprovincialis is a common aquaculture species, and also a major fouling organism that has negative economic impacts. There are no standard assay conditions for this important species and therefore, this study quantified the effect of key factors on the settlement of pediveligers and plantigrades. Density dependent settlement did not occur for either pediveligers or plantigrades. Settlement increased in drop assays in a 12 h light:12 h dark cycle, while bottom shade had no effect of any magnitude. In addition, settlement was significantly enhanced by storing pediveligers for between 4 and 24 days at 4 °C. Overall, these data provide the template to optimise and standardise static laboratory settlement assays for mussels in order to develop materials that either enhance settlement for the aquaculture industry, or deter settlement for antifouling applications. Furthermore, simple mechanisms such as storage at 4 °C can enhance settlement beyond current methods used in aquaculture hatcheries.