A deep chlorophyll layer (DCL) is a common feature of many deep, oligotrophic lakes including Lake Superior. Mechanisms generating and maintaining DCLs are variable across lakes, and seasonal patterns and relationships of DCL structure to physical variables are not well described. Using vertical profile data for physical and biological variables from western and central Lake Superior, we described seasonal patterns in DCL structure and other physical and biological parameters and applied linear mixed-effects models to determine how different physical factors (surface temperature, thermocline depth, and 1% photosynthetically active radiation (PAR) depth) affect the depth, thickness, maximum concentration, and integrated chlorophyll of the DCL. We observed clear seasonal patterns in the development and degradation of the DCL that coincide with seasonal changes in light and temperature. Modeling analysis using linear mixed-effects models showed that the DCL thickness was best predicted by surface temperature (R2 = 0.51) followed by thermocline depth (R2 = 0.36), and the deep chlorophyll maximum (DCM) concentration was best predicted by surface temperature (R2 = 0.26). The 1% PAR depth was not implicated as an important predictor, but observations from seasonal data suggest that it plays a role in the depth of the DCM. While no relationship was found between surface temperature and DCL-integrated chlorophyll, DCL thickness decreased and DCM concentration increased with increasing surface temperature, which could have implications for productivity in the DCL as the lake continues to warm.