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Assessment of metagenomic assembly using simulated next generation sequencing data

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Item Type:Article
Title:Assessment of metagenomic assembly using simulated next generation sequencing data
Creators Name:Mende, D.R. and Waller, A.S. and Sunagawa, S. and Jaervelin, A.I. and Chan, M.M. and Arumugam, M. and Raes, J. and Bork, P.
Abstract:Due to the complexity of the protocols and a limited knowledge of the nature of microbial communities, simulating metagenomic sequences plays an important role in testing the performance of existing tools and data analysis methods with metagenomic data. We developed metagenomic read simulators with platform-specific (Sanger, pyrosequencing, Illumina) base-error models, and simulated metagenomes of differing community complexities. We first evaluated the effect of rigorous quality control on Illumina data. Although quality filtering removed a large proportion of the data, it greatly improved the accuracy and contig lengths of resulting assemblies. We then compared the quality-trimmed Illumina assemblies to those from Sanger and pyrosequencing. For the simple community (10 genomes) all sequencing technologies assembled a similar amount and accurately represented the expected functional composition. For the more complex community (100 genomes) Illumina produced the best assemblies and more correctly resembled the expected functional composition. For the most complex community (400 genomes) there was very little assembly of reads from any sequencing technology. However, due to the longer read length the Sanger reads still represented the overall functional composition reasonably well. We further examined the effect of scaffolding of contigs using paired-end Illumina reads. It dramatically increased contig lengths of the simple community and yielded minor improvements to the more complex communities. Although the increase in contig length was accompanied by increased chimericity, it resulted in more complete genes and a better characterization of the functional repertoire. The metagenomic simulators developed for this research are freely available.
Keywords:Bacterial DNA, Bacterial Genome, Computational Biology, Computer Simulation, Contig Mapping, DNA Sequence Analysis, Genetic Models, Genomics, Metagenome, Metagenomics, Probability, Quality Control, Reproducibility of Results, Software
Source:PLoS ONE
ISSN:1932-6203
Publisher:Public Library of Science
Volume:7
Number:2
Page Range:e31386
Date:23 February 2012
Official Publication:https://doi.org/10.1371/journal.pone.0031386
PubMed:View item in PubMed

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