Mercury contamination of aquatic ecosystems is an issue of national concern, affecting both wildlife and human health. Detailed information on mercury cycling and food-web bioaccumulation in stream settings and the factors that control these processes is currently limited. In response, the U.S. Geological Survey (USGS) National Water-Quality Assessment Program (NAWQA) conducted detailed studies from 2002 to 2006 on various media to enhance process-level understanding of mercury contamination, biogeochemical cycling, and trophic transfer. Eight streams were sampled for this study: two streams in Oregon, and three streams each in Wisconsin and Florida. Streambed-sediment and pore-water samples were collected between February 2003 and September 2004. This report summarizes the suite of geochemical and microbial constituents measured, the analytical methods used, and provides the raw data in electronic form for both bed-sediment and pore-water media associated with this study.