We reconstructed late Holocene fluctuations of Kluane Lake in Yukon Territory from variations in bulk physical properties and carbon and nitrogen elemental and isotopic abundances in nine sediment cores. Fluctuations of Kluane Lake in the past were controlled by changes in climate and glaciers, which affected inflow of Slims and Duke rivers, the two largest sources of water flowing into the lake. Kluane Lake fluctuated within a narrow range, at levels about 25 m below the present datum, from about 5000 to 1300 cal yr BP. Low lake levels during this interval are probably due to southerly drainage of Kluane Lake to the Pacific Ocean, opposite the present northerly drainage to Bering Sea. Slims River, which today is the largest contributor of water to Kluane Lake, only rarely flowed into the lake during the period 5000 to 1300 cal yr BP. The lake rose 7–12 m between 1300 and 900 cal yr BP, reached its present level around AD 1650, and within a few decades had risen an additional 12 m. Shortly thereafter, the lake established a northern outlet and fell to near its present level.