Constructing a real metagenomic dataset with all known genomes for binning benchmarking. Organisms in the metagenome F1RT

So far, a lot of approaches have been developed to untangle metagenome-assembled sequences. For real metagenomic studies, binning performance is mainly assessed based on essential single-copy genes (SCGs) due to unknown of most genomes. However, SCGs only cover a small fraction of genomes and thereby just approximate binning performance. Here, we continued investigating the metagenome F1RT reported by us previously, considering high possibility to unearth all metagenome-assembled genomes (MAGs) due to its low complexity. Four isolates were obtained and found to fortunately separate remaining components in terms of sequencing coverage. Thus, an elaborate pipeline was in-house developed to obtain all MAGs. Strikingly, a novel MAGs termed FC8 was discovered here and 6 novel species were unearthed. A series of assessment extensively proofed its high reliability. Besides, our findings showed that this data set harbors several features, which are challenging for binning and thereby suitable to benchmark novel binning tools or compare advanced binning tools available now. Finally, using this data set, eight advanced binning algorithms were assessed, giving useful insights for developing novel approaches. To our knowledge, it is the first time to construct a real data set with all known MAGs. We hope that this data set will facilitate binning studies in future.