This data set is associated with an examination of environmental DNA (eDNA) from the invasive round goby (Neogobius melanostomus) in parallel with traditional fish surveys in the Great Lakes. Data resulting from bottom trawling surveys and angler hook and line methods include round goby fish count and biomass at different depths and seasons at multiple sites of Lake Huron and Lake Michigan (trawls), and breakwall locations of Lake Michigan at Portage Lakefront, Portage, Indiana and Washington Park, Michigan City, Indiana (angling). Data from eDNA water samples targeting round goby were analyzed by two molecular methods: droplet digital PCR (ddPCR) and quantitative PCR (qPCR); eDNA copy numbers are presented for both these methods separately.