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Ultra-deep, long-read nanopore sequencing of mock microbial community standards

2019· article· en· 341 citations· W2950671516 on OpenAlex· 10.1093/gigascience/giz043

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Canadian funderA Canadian agency funded it. The work may carry no Canadian affiliation at all.

No Canadian affiliation. An affiliation-only frame — the usual design — would never have seen this work. It is one of the works that make the case for inverting the frame.

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Opus teacher head0.012
GPT teacher head0.244
Teacher spread
0.232 · how far apart the two teachers sit on this one work
Validation status
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

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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The record

Venue
GigaScience
Topic
Genomics and Phylogenetic Studies
Field
Biochemistry, Genetics and Molecular Biology
Canadian institutions
Funders
University of British ColumbiaNortheastern UniversityMedical Research CouncilUniversity of BirminghamOntario Institute for Cancer ResearchNational Institute for Health and Care ResearchOxford Nanopore TechnologiesUniversity of NottinghamUniversity of QueenslandCornell University
Keywords
MetagenomicsNanopore sequencingDeep sequencingSequence assemblyComputational biologyMicrobial population biologyIllumina dye sequencingDNA sequencingBiologyNanoporeComputer scienceGenomeData miningGeneticsGeneNanotechnologyBacteria
Has abstract in OpenAlex
yes