PRJDA68519 The Usefulness and Reproducibility of Pyrosequencing for the Analysis of a Microbial Community of a Methane-Oxidizing Biofilm The usefulness of pyrosequencing for uncovering a methane-oxidizing biofilm community was determined in this study. Four individual DNA samples were prepared from a methanotrophic biofilm, and a multiplex pyrosequencing was performed. A complete library (sum of the 4 libraries) contained 33,639 sequences with an average length of 415 bp. It was interesting that methanotrophs were not dominant when grouped by functionality, making up only 23% of the community. Methylosinus, Methylomonas and Methylosarcina were the dominant methanotrophs. Type II methanotrophs were more abundant than type I (56 vs. 44%), but diversity and species richness of the type II community were lower. Dominant non-methanotrophic genera included Hydrogenophaga, Flavobacterium and Hyphomicrobium. For evaluation of the reproducibility, the complete library was de-multiplexed into 4 libraries, with different sequencing efforts, ranging from 3,915 to 20,133 sequences. Sørrenson abundance similarity results showed that the four libraries were almost identical (the indices > 0.97), and phylogenetic comparisons using UniFrac and P-tests revealed the same results. The 4 libraries had the same taxonomical composition, with similar relative abundances after taxonomic classification. EWHA_EBL_NRL study bioproject PRJDA68519 PRJDA68519 true genomeprj 68519 68519 false none provided