Carbon isotopes (14C and 13C) of dissolved organic carbon (DOC) in a Japanese oligotrophic lake (Lake Towada) were measured to study the origin and cycling of dissolved organic matter (DOM) in Lake Towada. Lake water samples were collected at 3 depths (0, 30, and 80 or 85 m) during 4 months (April, June, August, and October) in 2006. 14C measurements of DOC were performed by accelerator mass spectrometry (AMS) at the National Institute for Environmental Studies (NIES-TERRA) in Japan. Δ14C and δ13C values of DOC in Lake Towada showed light carbon isotopic values ranging from –750 to −514‰ and –29.0 to –27.8‰, respectively. These values are similar to those of humic substances reported. The very low carbon isotopic values of DOC in Lake Towada suggest a very small contribution of DOC derived from fresh phytoplankton to the lake DOC. There is an extremely high linear relationship between the Δ14C and δ13C of DOC in Lake Towada when all data points are plotted (r2 =0.818, p < 0.01), suggesting that the DOC in Lake Towada has 2 specific sources contributing heavy and light carbon isotopes. Although the freshly produced DOC of phytoplankton origin can be decomposed easily, the variation in the autochthonous DOC should influence the carbon isotopic values of DOC in Lake Towada.