Resolving complexity
Generating new insights into how microbial communities function requires improved approaches. Long-read sequencing enables species-level resolution and drives functional insights with full-length 16S rDNA and shotgun sequencing for metagenomic profiling and assemblies.
Single Molecule, Real-Time (SMRT) Sequencing for metagenomics pairs long reads with high accuracy, enabling you to:
- Understand key community functions by sequencing complete operons at 99.9% accuracy with long-insert metagenomic profiling – no assembly required
- Identify microbial community members with strain-level resolution using full-length 16S rDNA or shotgun sequencing
- Discover novel genes and gene clusters by reconstructing multi-kilobase long contigs with whole genome shotgun metagenomic assembly
- Leverage epigenomic data to associate contigs and plasmids from closely related strains
 | Library & Sample Preparation |
| Prepare targeted sequences or shotgun libraries to characterize your microbial community. |
 | Sequencing |
| Choose sequencing parameters to achieve 250-500K high-accuracy reads per SMRT Cell 1M on the Sequel System, depending on your library size. |
 | Data Analysis |
| Make discoveries using bioinformatics tools in SMRT Analysis or PacBio DevNet. |
Spotlight: Strain-level identification of bacterial communities with the unprecedented accuracy of PacBio full-length 16S sequencing
Using DADA2 software, all copies of the 16S housekeeping gene can be recovered from microbial communities. 16S sequence variants from the same strain appear in the data in integer ratios that reflect their copy number in each genome. In some cases, for example E. coli O157:H7 in the Zymo mock community, this high-resolution information can be used to unambiguously identify not only the species but the strain present. Explore this research further:
Callahan, B. J., et al. (2018). High-throughput amplicon sequencing of the full-length 16S rRNA gene with single-nucleotide resolution. BioRxiv, Preprint.
Long reads and methylation profiles enable binning of metagenome assembly contigs and plasmids
SMRT Sequencing of metagenomes enables highly contiguous assemblies of community members, but leveraging the associated methylation information further allows those contigs to be associated into near-complete genome bins. Plasmids of sufficient size can also be associated with hosts via their methylation fingerprints. Explore this research further:
Beaulaurier, J., et al. (2017). Metagenomic binning and association of plasmids with bacterial host genomes using DNA methylation. Nature Biotechnology, 36(1), 61–69.
Selected Resources
- Nilsson, R Henrik et al. (2018) Mycobiome diversity: high-throughput sequencing and identification of fungi. Nature reviews. Microbiology
- Yan, Bo et al. (2018) SMRT-Cappable-seq reveals complex operon variants in bacteria. Nature communications
- Callahan, Benjamin J et al. (2018) High-throughput amplicon sequencing of the full-length 16S rRNA gene with single-nucleotide resolution BioRxiv
- Hiraoka, Satoshi et al. (2018) Metaepigenomic analysis reveals the unexplored diversity of DNA methylations in an environmental prokaryotic community BioRxiv
- Redondo, Miguel A et al. (2018) Contrasting distribution patterns between aquatic and terrestrial Phytophthora species along a climatic gradient are linked to functional traits. The ISME journal
- Heeger, Felix et al. (2018) Long-read DNA metabarcoding of ribosomal RNA in the analysis of fungi from aquatic environments. Molecular ecology resources
- P.E. Zida et al. (2018) Increasing sorghum yields by seed treatment with an aqueous extract of the plant Eclipta alba may involve a dual mechanism of hydropriming and suppression of fungal pathogens Crop protection
- Tedersoo, Leho et al. (2018) PacBio metabarcoding of Fungi and other eukaryotes: errors, biases and perspectives. The New phytologist
- Beaulaurier, John et al. (2017) Metagenomic binning and association of plasmids with bacterial host genomes using DNA methylation. Nature biotechnology
- Perraudeau, Fanny et al. (2017) Accurate determination of bacterial abundances in human metagenomes using full-length 16S sequencing reads BioRxiv
- Wang, Yuan et al. (2017) Profiling of oral microbiota in early childhood caries using Single-Molecule Real-Time Sequencing Frontiers in microbiology
- Edwards, Joan E et al. (2017) PCR and omics based techniques to study the diversity, ecology and biology of anaerobic fungi: Insights, challenges andopportunities. Frontiers in microbiology
- Zhao, J et al. (2017) Reduction in fecal microbiota diversity and short-chain fatty acid producers in Methicillin-resistant Staphylococcus aureus infected individuals as revealed by PacBio single molecule, real-time sequencing technology. European journal of clinical microbiology & infectious diseases
- Nakano, Kazuma et al. (2017) Advantages of genome sequencing by long-read sequencer using SMRT technology in medical area. Human Cell
- Tai, Phillip WL et al. (2018) Adeno-associated virus genome population sequencing achieves full vector genome resolution and reveals human-vector chimeras Molecular therapy
- Xu, Haiyan et al. (2017) The effects of probiotics administration on the milk production, milk components and fecal bacteria microbiota of dairy cows Science bulletin
- Pootakham, Wirulda et al. (2017) High resolution profiling of coral-associated bacterial communities using full-length 16S rRNA sequence data from PacBio SMRT sequencing system. Scientific reports
- Sadowsky, Michael J et al. (2017) Analysis of gut microbiota – An ever changing landscape. Gut microbes
- McRose, Darcy L et al. (2017) Diversity and activity of alternative nitrogenases in sequenced genomes and coastal environments. Frontiers in microbiology
- Meng, Xiangli et al. (2017) Metataxonomics reveal vultures as a reservoir for Clostridium perfringens. Emerging microbes and infections
- Motooka, Daisuke et al. (2017) Fungal ITS1 deep-sequencing strategies to reconstruct the composition of a 26-species community and evaluation of the gut mycobiota of healthy Japanese individuals. Frontiers in microbiology
- Driscoll, Connor B et al. (2017) Towards long-read metagenomics: complete assembly of three novel genomes from bacteria dependent on a diazotrophic cyanobacterium in a freshwater lake co-culture. Standards in genomic sciences
- Hagen, Live H et al. (2017) Quantitative metaproteomics highlight the metabolic contributions of uncultured phylotypes in a thermophilic anaerobic digester. Applied and environmental microbiology
- Li, Jing et al. (2017) Bacterial microbiota of Kazakhstan cheese revealed by single molecule real time (SMRT) sequencing and its comparison with Belgian, Kalmykian and Italian artisanal cheeses BMC microbiology
- Chen, Yuan et al. (2015) Next generation multilocus sequence typing (NGMLST) and the analytical software program MLSTEZ enable efficient, cost-effective, high-throughput, multilocus sequencing typing. Fungal genetics and biology
- Poster: Boitano, M. et al. (2018) Full-length cDNA sequencing of prokaryotic transcriptome and metatranscriptome samples
- Poster: Hall, Richard et al. (2017) Using the PacBio Sequel System to taxonomically and functionally classify metagenomic samples in a trial of patients undergoing fecal microbiota transplantation
- Poster: Heiner, C. et al. (2017) Profiling complex population genomes with highly accurate single molecule reads: cow rumen microbiomes
- Poster: Oh, S. et al. (2016) Minimization of chimera formation and substitution errors in full-length 16S PCR amplification
- Poster: Heiner, C. et al. (2016) WGS SMRT Sequencing of patient samples from a fecal microbiota transplant trial
- Poster: Hall, R. et al. (2016) Workflow for processing high-throughput, Single Molecule, Real-Time Sequencing data for analyzing the microbiome of patients undergoing fecal microbiota transplantation
- Poster: Heiner, Cheryl et al. (2016) Low-input long-read sequencing for complete microbial genomes and metagenomic community analysis
- Poster: Hall, Richard et al. (2016) Profiling the microbiome in fecal microbiota transplantation using circular consensus and Single Molecule, Real-Time Sequencing
- Poster: Hall, Richard J et al. (2016) Profiling metagenomic communities using circular consensus and Single Molecule, Real-Time Sequencing
- Ehrlich, Garth (2018) User Group Meeting: Development, validation, and applications of a high-fidelity, species-specific, eubacterial pan-domain microbiome assay on the PacBio System
- Hatas, Emily (2017) Webinar: SMRT Sequencing applications in plant and animal sciences: an overview
- Auvignen, Petra (2017) AGBT Virtual Poster: Interspecies interation amoung meat spoilage-related lactic acid bacteria
- (2017) Tutorial: Circular Consensus Sequence analysis application [SMRT Link v5.0.0]
- Heiner, Cheryl (2016) AGBT Virtual Poster: Low-input long-read sequencing for complete microbial genomes and metagenomic community analysis
- Heiner, Cheryl (2016) AGBT Virtual Poster: Minimization of chimera formation and substitution errors in full-length 16S PCR amplification
- Hall, Richard (2016) AGBT Virtual Poster: Profiling the microbiome in fecal microbiota transplantation using circular consensus and Single Molecule, Real-Time Sequencing
- PacBio Certified Service Providers (2018)
- SMRT Sequencing Brochure: Revolutionize genomics with SMRT Sequencing (2018)
- Application Brochure: Microbial communities – Complex microbial populations simplified. (2017)
- SMRT Analysis Brochure: Gain a deeper understanding of your sequencing data (2017)
- Industrial Biotechnology Brochure: Fuel biotech discovery with confident characterization of microbes and their communities. (2016)
- Epigenetics Application Brochure: Characterize the epigenetic landscape of your genome. (2016)