Abstract

The development of high-throughput sequencing methods allows for the characterization of microbial communities in a wide range of environments on an unprecedented scale. However, insight into microbial community composition is limited by our ability to detect patterns in this flood of sequences. Here we compare the performance of 51 analysis techniques using real and simulated bacterial 16S rRNA pyrosequencing datasets containing either clustered samples or samples arrayed across environmental gradients. We find that many diversity patterns are evident with severely undersampled communities, and that methods vary widely in their ability to detect gradients and clusters. Chi-squared distances and Pearson correlation distances perform especially well for detecting gradients, while Gower and Canberra distances perform especially well for detecting clusters. These results also provide a basis for understanding tradeoffs between number of samples and depth of coverage, tradeoffs which are important to consider when designing studies to characterize microbial communities.

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