Permafrost environments within the Siberian Arctic are natural sources of the climate relevant trace gas methane. In order to improve our understanding of the present and future carbon dynamics in high latitudes, we studied the methane concentration, the quantity and quality of organic matter, and the activity and biomass of the methanogenic community in permafrost deposits. For these investigations a permafrost core of Holocene age was drilled in the Lena Delta (72°22′N, 126°28′E). The organic carbon of the permafrost sediments varied between 0.6% and 4.9% and was characterized by an increasing humification index with permafrost depth. A high CH4 concentration was found in the upper 4 m of the deposits, which correlates well with the methanogenic activity and archaeal biomass (expressed as PLEL concentration). Even the incubation of core material at −3 and −6°C with and without substrates showed a significant CH4 production (range: 0.04–0.78 nmol CH4 h−1 g−1). The results indicated that the methane in Holocene permafrost deposits of the Lena Delta originated from modern methanogenesis by cold-adapted methanogenic archaea. Microbial generated methane in permafrost sediments is so far an underestimated factor for the future climate development.