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Microfluidic-based mini-metagenomics enables discovery of novel microbial lineages from complex environmental samples

View ORCID ProfileFeiqiao Brian Yu, View ORCID ProfilePaul C. Blainey, View ORCID ProfileFrederik Schulz, Tanja Woyke, Mark A. Horowitz, Stephen R. Quake
doi: https://doi.org/10.1101/114496
Feiqiao Brian Yu
1Department of Electrical Engineering, Stanford University, 350 Serra Mall, Stanford, CA 94305
2Department of Bioengineering, Stanford University, 443 Via Ortega, Stanford, CA 94305
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Paul C. Blainey
3MIT Department of Biological Engineering and Broad Institute of Harvard and MIT, 415 Main Street, Cambridge, MA 02142
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Frederik Schulz
4Department of Energy Joint Genome Institute, 2800 Mitchell Drive, Walnut Creek, CA 94598
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Tanja Woyke
4Department of Energy Joint Genome Institute, 2800 Mitchell Drive, Walnut Creek, CA 94598
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Mark A. Horowitz
1Department of Electrical Engineering, Stanford University, 350 Serra Mall, Stanford, CA 94305
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Stephen R. Quake
2Department of Bioengineering, Stanford University, 443 Via Ortega, Stanford, CA 94305
5Chan Zuckerberg Biohub, San Francisco, CA 94158, USA
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  • For correspondence: quake@stanford.edu
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Abstract

Metagenomics and single-cell genomics have enabled the discovery of many new genomes from previously unknown branches of life. However, extracting novel genomes from complex mixtures of metagenomic data can still be challenging and in many respects represents an ill-posed problem which is generally approached with ad hoc methods. Here we present a microfluidic-based mini-metagenomic method which offers a statistically rigorous approach to extract novel microbial genomes from complex samples. In addition, by generating 96 sub-samples from each environmental sample, this method maintains high throughput, reduces sample complexity, and preserves single-cell resolution. We used this approach to analyze two hot spring samples from Yellowstone National Park and extracted 29 new genomes larger than 0.5 Mbps. These genomes represent novel lineages at different taxonomic levels, including three deeply branching lineages. Functional analysis revealed that these organisms utilize diverse pathways for energy metabolism. The resolution of this mini-metagenomic method enabled accurate quantification of genome abundance, even for genomes less than 1% in relative abundance. Our analyses also revealed a wide range of genome level single nucleotide polymorphism (SNP) distributions with nonsynonymous to synonymous ratio indicative of low to moderate environmental selection. The scale, resolution, and statistical power of microfluidic-based mini-metagenomic make it a powerful tool to dissect the genomic structure microbial communities while effectively preserving the fundamental unit of biology, the single cell.

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Posted March 07, 2017.
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Microfluidic-based mini-metagenomics enables discovery of novel microbial lineages from complex environmental samples
Feiqiao Brian Yu, Paul C. Blainey, Frederik Schulz, Tanja Woyke, Mark A. Horowitz, Stephen R. Quake
bioRxiv 114496; doi: https://doi.org/10.1101/114496
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Microfluidic-based mini-metagenomics enables discovery of novel microbial lineages from complex environmental samples
Feiqiao Brian Yu, Paul C. Blainey, Frederik Schulz, Tanja Woyke, Mark A. Horowitz, Stephen R. Quake
bioRxiv 114496; doi: https://doi.org/10.1101/114496

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