INVIEW Microbiome 16S-Full-Length

INVIEW Microbiome 16S Full-Length
Nanopore Sequencing

INVIEW Microbiome profiling using long reads

Microbiome profiling using long reads leverages third-generation sequencing technologies like Oxford Nanopore to sequence longer DNA fragments, such as the full-length ~1.5 kb 16S rRNA gene, enabling higher taxonomic resolution. This method offers greater accuracy in identifying and classifying microorganisms down to the species and strain level, a challenge with short-read sequencing.

By capturing full-length sequences, long-read profiling enhances the ability to distinguish closely related microbial species, providing a more detailed and comprehensive view of microbial diversity.

If you prepare your own amplicons, we recommend utilizing our short- and long-read amplicon sequencing services.

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Applications

Characterization of strains or communities
Identification of taxonomic background
Metagenomic analysis of long reads

Highlights

Full-length amplification of ~1.5 kb 16S rRNA gene using V1-V9 primers (27 F and 1492R)
Ligation based library preparation
Sequencing using Oxford Nanopore GridION (flow cell R10.4.1)
Pooling of up to 96 samples per flow cell
Data transfer via secure FTP
Services are performed in ISO17025 certified labs

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Product Specifications & Ordering

Pre-sequencing options

Optional DNA isolation service: from various tissue types, cell lines, fermented products, cheese or yoghurt, enrichment cultures / starter cultures, buccal swabs, human and animal feces, water & waste water, earthy soil samples (no sand). Please refer to our DNA isolation guide.

Please note that we do not accept samples with higher biosafety level than S2. GMOs are only accepted with S1 level.

Starting material

Sample Type: Purified DNA
Purity (OD260/280): 1.8-2.0
Volume: 20µL
Concentration: 10-50 ng/µl
Minimum Amount: 200 ng
Resuspension Buffer Water, EB, or low TE (<0.1 mM EDTA)
Format: barcode labelled 1.5 ml safe-lock tubes; or for > 48 samples in Eppendorf twin.tec PCR Plate 96, full-skirted, leave position G12 & H12 empty
Shipment Method: Room temperature/ice

Product description

Library preparation

Full-length amplification of ~1.5 kb 16S rRNA gene using V1-V9 primers (27F and 1492R)
Ligation-based, fragmentation-free library preparation (amplification-free) using v14 library prep chemistry
Pooling of up to 96 samples per flow cell

Sequencing

Technology: Oxford Nanopore GridION (flow cell R10.4.1)
Output read length: Equal to fragment length (for full-length 16S rRNA gene approx. 1500 bp)
Output varies based on the number of amplicons, with approximately 50,000 reads per amplicon typically achieved for robust data generation.

Deliverables

Raw Nanopore sequencing data (FastQ)

We provide optional bioinformatics for comprehensive 16S microbiome profiling

Bioinformatics analysis

Rawdata FastQ Files (sequences and quality scores)

Optional analysis for comprehensive microbiome profiling:

Additional FASTQ files
- Sequence cleaned
- Classified
- Unclassified
Taxonomical assignment and read abundance estimation for all detected OTUs down to species level
Comprehensive profiling report (.HTML)
Pipeline read statistics including classification rate
Abundance table with counts per taxa
The following delivered at different taxonomic rank levels
- Bar plot comparing relative abundances of high abundant taxa in all samples
- Alpha diversity metrices (diversity within samples)
  - Shannon index
  - Simpson index
  - etc.
- Beta diversity analyses (diversity between samples)
  - Principal coordinates analysis (PCoA)
  - Distance metric plus heatmap