Comparative Analysis of Functional Metagenomic Annotation and the Mappability of Short Reads
To assess the functional capacities of microbial communities, including those inhabiting the human body, shotgun metagenomic reads are often aligned to a database of known genes. Such homology-based annotation practices critically rely on the assumption that short reads can map to orthologous genes...
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| Format: | Online |
| Language: | English |
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Public Library of Science
2014
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| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4141809/ |
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pubmed-4141809 |
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pubmed-41418092014-08-25 Comparative Analysis of Functional Metagenomic Annotation and the Mappability of Short Reads Carr, Rogan Borenstein, Elhanan Research Article To assess the functional capacities of microbial communities, including those inhabiting the human body, shotgun metagenomic reads are often aligned to a database of known genes. Such homology-based annotation practices critically rely on the assumption that short reads can map to orthologous genes of similar function. This assumption, however, and the various factors that impact short read annotation, have not been systematically evaluated. To address this challenge, we generated an extremely large database of simulated reads (totaling 15.9 Gb), spanning over 500,000 microbial genes and 170 curated genomes and including, for many genomes, every possible read of a given length. We annotated each read using common metagenomic protocols, fully characterizing the effect of read length, sequencing error, phylogeny, database coverage, and mapping parameters. We additionally rigorously quantified gene-, genome-, and protocol-specific annotation biases. Overall, our findings provide a first comprehensive evaluation of the capabilities and limitations of functional metagenomic annotation, providing crucial goal-specific best-practice guidelines to inform future metagenomic research. Public Library of Science 2014-08-22 /pmc/articles/PMC4141809/ /pubmed/25148512 http://dx.doi.org/10.1371/journal.pone.0105776 Text en © 2014 Carr, Borenstein http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
| repository_type |
Open Access Journal |
| institution_category |
Foreign Institution |
| institution |
US National Center for Biotechnology Information |
| building |
NCBI PubMed |
| collection |
Online Access |
| language |
English |
| format |
Online |
| author |
Carr, Rogan Borenstein, Elhanan |
| spellingShingle |
Carr, Rogan Borenstein, Elhanan Comparative Analysis of Functional Metagenomic Annotation and the Mappability of Short Reads |
| author_facet |
Carr, Rogan Borenstein, Elhanan |
| author_sort |
Carr, Rogan |
| title |
Comparative Analysis of Functional Metagenomic Annotation and the Mappability of Short Reads |
| title_short |
Comparative Analysis of Functional Metagenomic Annotation and the Mappability of Short Reads |
| title_full |
Comparative Analysis of Functional Metagenomic Annotation and the Mappability of Short Reads |
| title_fullStr |
Comparative Analysis of Functional Metagenomic Annotation and the Mappability of Short Reads |
| title_full_unstemmed |
Comparative Analysis of Functional Metagenomic Annotation and the Mappability of Short Reads |
| title_sort |
comparative analysis of functional metagenomic annotation and the mappability of short reads |
| description |
To assess the functional capacities of microbial communities, including those inhabiting the human body, shotgun metagenomic reads are often aligned to a database of known genes. Such homology-based annotation practices critically rely on the assumption that short reads can map to orthologous genes of similar function. This assumption, however, and the various factors that impact short read annotation, have not been systematically evaluated. To address this challenge, we generated an extremely large database of simulated reads (totaling 15.9 Gb), spanning over 500,000 microbial genes and 170 curated genomes and including, for many genomes, every possible read of a given length. We annotated each read using common metagenomic protocols, fully characterizing the effect of read length, sequencing error, phylogeny, database coverage, and mapping parameters. We additionally rigorously quantified gene-, genome-, and protocol-specific annotation biases. Overall, our findings provide a first comprehensive evaluation of the capabilities and limitations of functional metagenomic annotation, providing crucial goal-specific best-practice guidelines to inform future metagenomic research. |
| publisher |
Public Library of Science |
| publishDate |
2014 |
| url |
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4141809/ |
| _version_ |
1613126810711621632 |