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Average genome size (AGS) is an important, yet often overlooked property of microbial communities. We developed MicrobeCensus to rapidly and accurately estimate AGS from short-read metagenomics data and applied our tool to over 1,300 human microbiome samples. We found that AGS differs significantly within and between body sites and tracks with major functional and taxonomic differences. For example, in the gut, AGS ranges from 2.5 to 5.8 megabases and is positively correlated with the abundance of Bacteroides and polysaccharide metabolism. Furthermore, we found that AGS variation can bias comparative analyses, and that normalization improves detection of differentially abundant genes.

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