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We used phospholipid fatty acids (PLFA) to determine microbial biomass and community structure in the sediments of eight boreal lakes with different loadings of allochthonous organic carbon and total phosphorus (TP) in the water during the course of a year. The total concentration of PLFA, an estimate of the microbial biomass, depended more on TP, a proxy for pelagic primary production, but not on dissolved organic carbon, a proxy for terrestrial organic carbon input. The composition of PLFAs varied considerably over time, demonstrating seasonal dynamics in microbial community composition. When PLFA profiles in all lakes and seasons are compared, community composition is more similar within season than within lakes.