The main river inflows to Loch Ness and several depths in the water column within the loch were sampled over an annual cycle. The carbon isotope composition of total dissolved inorganic carbon (DIC) from the samples was determined as well as that of phytoplankton from the loch. Values of δ13C for DIC in the rivers indicated that this DIC was derived from soil respiration in the catchment and achieved only partial equilibrium with the atmosphere during river transport. Riverine loading accounted for most of the DIC in Loch Ness, and the great depth of the loch relative to its surface area allows only limited exchange with the atmosphere. Despite the low productivity in Loch Ness, DIC concentrations in the low alkalinity water are appreciably influenced by plankton metabolism, and seasonal fluctuations in δ13C of DIC and phytoplankton revealed the particular impact of photosynthetic carbon fixation on DIC. However, the photosynthetic depletion of DIC during summer does not offset the riverine loading sufficiently to prevent the loch waters being supersaturated with CO2 throughout the year. Annual efflux of CO2 from Loch Ness is estimated to be 253 × 106 mol yr−1, of which around one quarter may be due to net heterotrophic mineralization within the loch of organic carbon of terrestrial origin. The remainder is attributable to inorganic carbon input to the lake via river inflow and derived from prior mineralization of soil organic matter within the drainage area. This annual efflux of CO2 can represent around 6% of net ecosystem production in the catchment.