These data pertain to water and sediments collected from the Saint Louis River Estuary (SLRE) and its nearby water sources by Luke Loken and collaborators for his Masters thesis and additional publications. In brief, we sampled SLRE surface waters and sediments for a variety of physical, chemical, and biological attributes. Ten estuary stations were sampled approximately monthly from April 2012 through September 2013. On four of the sampling campaigns, water was collected from an additional 20 sites. Sites were selected to represent a gradient from the Saint Louis River to Lake Superior and included several tributaries that drain directly into the estuary. This design aimed to understand the spatial and temporal mixing pattern of the estuary as it receives water from several rivers, 2 waste water treatment plant, and Lake Superior. We sampled the estuary to assess the magnitude and timing of source water contributions to the estuary and establish a baseline of chemical and physical measurements to aid in future limnological research. Additionally, we performed nitrogen and carbon cycling rate experiments to determine the estuary-wide influence on nitrate, ammonium, and dissolved organic carbon. This included 8 sediment denitrification, 1 nitrification, and 2 breakdown dissolved organic carbon (BDOD) surveys. This work was funded by the Minnesota and Wisconsin Sea Grant and in coordination with the establishment of the Lake Superior National Estuary Research Reserve (LSNERR).