Microbiome Sequencing Solution

The easy end-to-end workflow provides significant improvement over many of the standard protocols that are known to miss microbes or provide biased results

PCR Sequencing Targets

Based on your experimental needs and the availability of sequencing platforms, Intus kits offer optimized PCR choices for parallel sequencing of over 100s to 1000s of samples leveraging the entire 16S region or the 16S gene plus the highly variable internally transcribed  spacer region depending on the taxa classification needs of the experiment.

The kits take the guesswork out of which target sequences to choose, what polymerase to use, and the level of primer degeneracy. They are designed and validated with multiplex primers for sequencing many samples simultaneously in a single sequencing run. The PCR conditions are optimized for sequencing a simple or a complex microbiome community and ensure conditions that result in correct microbial profiles.

The assay kits for the PacBio Sequel system contain prearrayed PCR wells for lysis, DNA purification beads, and prearrayed PCR wells containing dual-ended  barcoded primers targeting either the entire 16S rRNA gene or the extended 16S-ITS-23S gene region. Following PCR amplification, amplicons are pooled and ready for the addition of PacBio SMRTbell adaptors followed by sequencing on the PacBio Sequel System. Utilization of barcoded SMRT bell adaptors also allow further multiplexing of 100s to 1000s of samples on a single SMRT cell.

The assay kits for the Oxford Nanopore Technologies platform contain prearrayed PCR wells for lysis, DNA purification beads, and prearrayed PCR wells containing dual ended barcoded primers targeting the extended 16S-ITS-23S gene region. Following PCR amplification, amplicons are pooled and ready for Oxford Nanopore ligation adaptors followed by sequencing. 

2 Kit Configurations Available

• Intus Complete kits include the lysis, DNA purification, and PCR reagents needed to easily and quickly lyse and prepare amplicons for sequencing.
• Intus Wave kits provide the same PCR reagents for customers with already prepared high-quality DNA.

Data Analysis

The microbiome sequencing kits are only available when purchased together with a bioinformatics analysis service provided by Intus Biosciences. You can simply share your raw sequencing data with Intus to be analyzed using their microbiome analysis pipeline and proprietary reference database. The software is compatible with long-read amplicon data generated either from the PacBio or Oxford Nanopore sequencers, sorts samples by barcode and allows tracking by user-selected sample names. Each sample’s reads are mapped using the Athena Microbial Reference Database to determine phylogenetic identity.

Output can be transferred to a spreadsheet to compare results between individual samples or combine samples into virtual groups for comparison at the phylum, strain, or other taxonomic levels.

Mapping allows investigators to quickly test hypotheses about variations in microbial types between microbiome cases and controls. Reads can also be used for custom analysis, as desired.

The data analysis will be tailored to your specific needs. Therefore, a setup fee is due for your first purchase of an Intus Microbiome Service. Please contact us for a quotation.