Immense amounts of ancient (radiocarbon age over 200years) organic carbon (OC) from permafrost are released into aquatic systems. Ancient terrestrial OC exists in numerous aquatic ecosystems. It has been reported that ancient OC can be incorporated by consumers in aquatic ecosystems, but the effect of ancient OC on the growth of consumers has rarely been studied. In this study, we extracted ancient dissolved organic carbon (DOC) from frozen soils in an alpine lake catchment. After a 6-day microbial conversion period, the contents of ω3 and ω6 polyunsaturated fatty acids (PUFAs) in ancient DOC increased. Proteobacteria and Actinobacteria were the primary taxa consuming the permafrost DOC and generating fatty acids. In addition to the exclusive diet of soil DOC (containing bacteria) or Chlorella pyrenoidosa, mixed diets of Chlorella pyrenoidosa, and ancient DOC (containing bacteria) in ratios of 2:1, 1:1, and 1:2 (by carbon concentration) were used to feed Daphnia magna. We discovered that Daphnia reared on the mixture with the DOC:Chlorella ratio of 1:2 had the highest contents of ω3 PUFAs and FAs. Daphnia reared exclusively on Chlorella and the mixture with the DOC:Chlorella ratio of 1:2 had the largest body size (3.1-3.4mm) and the highest offspring production (95.5-96.2 ind-1). Daphnia fed on mixed diets exhibited higher intrinsic rates of population growth (0.48-0.53 d-1) compared to those fed exclusively on Chlorella pyrenoidosa, or ancient DOC plus bacteria. Overall, ancient soil OC converted by bacteria can act as a valuable supplement to algae food to promote Daphnia growth.