Ultra-deep, long-read nanopore sequencing of mock microbial community standards
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Abstract
Background
Long sequencing reads are information-rich: aiding de novo assembly and reference mapping, and consequently have great potential for the study of microbial communities. However, the best approaches for analysis of long-read metagenomic data are unknown. Additionally, rigorous evaluation of bioinformatics tools is hindered by a lack of long-read data from validated samples with known composition.
Findings
We sequenced 2 commercially available mock communities containing 10 microbial species (ZymoBIOMICS Microbial Community Standards) with Oxford Nanopore GridION and PromethION. Both communities and the 10 individual species isolates were also sequenced with Illumina technology. We generated 14 and 16 gigabase pairs from 2 GridION flowcells and 150 and 153 gigabase pairs from 2 PromethION flowcells for the evenly distributed and log-distributed communities, respectively. Read length N50 ranged between 5.3 and 5.4 kilobase pairs over the 4 sequencing runs. Basecalls and corresponding signal data are made available (4.2 TB in total). Alignment to Illumina-sequenced isolates demonstrated the expected microbial species at anticipated abundances, with the limit of detection for the lowest abundance species below 50 cells (GridION). De novo assembly of metagenomes recovered long contiguous sequences without the need for pre-processing techniques such as binning.
Conclusions
We present ultra-deep, long-read nanopore datasets from a well-defined mock community. These datasets will be useful for those developing bioinformatics methods for long-read metagenomics and for the validation and comparison of current laboratory and software pipelines.
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Now published in GigaScience doi: 10.1093/gigascience/giz043
Samuel M. Nicholls 1Institute of Microbiology and Infection, School of Biosciences, University of Birmingham, UKFind this author on Google ScholarFind this author on PubMedSearch for this author on this siteORCID record for Samuel M. NichollsJoshua C. Quick 1Institute of Microbiology and Infection, School of Biosciences, University of Birmingham, UKFind this author on Google ScholarFind this author on PubMedSearch for this author on this siteORCID record for Joshua C. QuickShuiquan Tang 2Zymo Research Corporation, Irvine, California, USAFind this author on Google ScholarFind this author on PubMedSearch for this author on this siteNicholas J. Loman 1Institute of Microbiology and Infection, School of Biosciences, University of Birmingham, UKFind this author on Google …
Now published in GigaScience doi: 10.1093/gigascience/giz043
Samuel M. Nicholls 1Institute of Microbiology and Infection, School of Biosciences, University of Birmingham, UKFind this author on Google ScholarFind this author on PubMedSearch for this author on this siteORCID record for Samuel M. NichollsJoshua C. Quick 1Institute of Microbiology and Infection, School of Biosciences, University of Birmingham, UKFind this author on Google ScholarFind this author on PubMedSearch for this author on this siteORCID record for Joshua C. QuickShuiquan Tang 2Zymo Research Corporation, Irvine, California, USAFind this author on Google ScholarFind this author on PubMedSearch for this author on this siteNicholas J. Loman 1Institute of Microbiology and Infection, School of Biosciences, University of Birmingham, UKFind this author on Google ScholarFind this author on PubMedSearch for this author on this siteORCID record for Nicholas J. Loman
A version of this preprint has been published in the Open Access journal GigaScience (see paper https://doi.org/10.1093/gigascience/giz043 ), where the paper and peer reviews are published openly under a CC-BY 4.0 license.
These peer reviews were as follows:
Reviewer 1: http://dx.doi.org/10.5524/REVIEW.101702 Reviewer 2: http://dx.doi.org/10.5524/REVIEW.101703
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