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Libra: scalable k-mer-based tool for massive all-vs-all metagenome comparisons
Background Shotgun metagenomics provides powerful insights into microbial community biodiversity and function. Yet, inferences from metagenomic studies are often limited by dataset size and complexity and are restricted by the availability and completeness of existing databases. De novo comparative metagenomics enables the comparison of metagenomes based on their total genetic content. Results We developed a tool called Libra that performs an all-vs-all comparison of metagenomes for precise clustering based on their k-mer content. Libra uses a scalable Hadoop framework for massive metagenome comparisons, Cosine Similarity for calculating the distance using sequence composition and abundance while normalizing for sequencing depth, and a web-based implementation in iMicrobe (http://imicrobe.us) that uses the CyVerse advanced cyberinfrastructure to promote broad use of the tool by the scientific community. Conclusions A comparison of Libra to equivalent tools using both simulated and real metagenomic datasets, ranging from 80 million to 4.2 billion reads, reveals that methods commonly implemented to reduce compute time for large datasets, such as data reduction, read count normalization, and presence/absence distance metrics, greatly diminish the resolution of large-scale comparative analyses. In contrast, Libra uses all of the reads to calculate k-mer abundance in a Hadoop architecture that can scale to any size dataset to enable global-scale analyses and link microbial signatures to biological processes.National Science Foundation [1640775]Open access journalThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]
Spaced seeds improve k-mer-based metagenomic classification
Metagenomics is a powerful approach to study genetic content of environmental
samples that has been strongly promoted by NGS technologies. To cope with
massive data involved in modern metagenomic projects, recent tools [4, 39] rely
on the analysis of k-mers shared between the read to be classified and sampled
reference genomes. Within this general framework, we show in this work that
spaced seeds provide a significant improvement of classification accuracy as
opposed to traditional contiguous k-mers. We support this thesis through a
series a different computational experiments, including simulations of
large-scale metagenomic projects. Scripts and programs used in this study, as
well as supplementary material, are available from
http://github.com/gregorykucherov/spaced-seeds-for-metagenomics.Comment: 23 page
Picoeukaryotic sequences in the Sargasso Sea metagenome
Many sequences from picoeukaryotes were found in DNA sequence data assembled from Sargasso seawater
Multiple Comparative Metagenomics using Multiset k-mer Counting
Background. Large scale metagenomic projects aim to extract biodiversity
knowledge between different environmental conditions. Current methods for
comparing microbial communities face important limitations. Those based on
taxonomical or functional assignation rely on a small subset of the sequences
that can be associated to known organisms. On the other hand, de novo methods,
that compare the whole sets of sequences, either do not scale up on ambitious
metagenomic projects or do not provide precise and exhaustive results.
Methods. These limitations motivated the development of a new de novo
metagenomic comparative method, called Simka. This method computes a large
collection of standard ecological distances by replacing species counts by
k-mer counts. Simka scales-up today's metagenomic projects thanks to a new
parallel k-mer counting strategy on multiple datasets.
Results. Experiments on public Human Microbiome Project datasets demonstrate
that Simka captures the essential underlying biological structure. Simka was
able to compute in a few hours both qualitative and quantitative ecological
distances on hundreds of metagenomic samples (690 samples, 32 billions of
reads). We also demonstrate that analyzing metagenomes at the k-mer level is
highly correlated with extremely precise de novo comparison techniques which
rely on all-versus-all sequences alignment strategy or which are based on
taxonomic profiling
The MGX framework for microbial community analysis
Jaenicke S. The MGX framework for microbial community analysis. Bielefeld: Universität Bielefeld; 2020
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