We examined the radiocarbon (14C) reservoir effect in Lake Kutubu using tephrochronology and terrestrial plant material to deliver a precise age-depth profile and sedimentation rates for this lake. Based on the presence of two tephra horizons (Tibito and Olgaboli), we found a reservoir age offset in sediments of between 1490 and 2280 14C yr using the sediment ages derived from the lead-210 (210Pb) dating method. The live submerged biological samples collected exhibited a higher reservoir age offset than the sediment. This is most likely a result of delayed transport of “bomb” 14C from the atmosphere to aquatic and sedimentary system. The 14C reservoir effect increased with distance from the lake inlet and also decreased with depth. Dissolution of 14C-depleted carbon from surrounding limestone and direct in-wash of old soil or vegetation remnants from the catchment are the most likely causes of the 14C reservoir effect. Based on limestone areas mapped in Papua New Guinea, we indicate lakes which may be subject to a significant 14C reservoir effect. The results of this study demonstrate the magnitude of the 14C reservoir effect in lakes and provide insights to the correct interpretation of past environmental and archaeological events in PNG.