Introduction to 16S rDNA Sequencing
The 16S rRNA gene is ubiquitous in prokaryotic organisms and is commonly utilized as a biomarker to differentiate between microbial phylogenies. Using 16S microbiome analysis, it is possible to get an overview of the community composition of a microbiome and identify differences between groups and variation within groups for fast and affordable investigation of any microbial community.
Sequencing of the 16S rRNA Gene
The whole 16S rRNA gene is approximately 1550 base pairs long and consists of nine variable regions (V1-V9) flanked by conserved regions critical to ribosomal functions. This structural setting can be utilized as variable regions are suitable as phylogenetic markers to differentiate between species, while the conserved regions serve as binding sites for primers that are designed to amplify the variable regions of interest using PCR.
The primers usually consist of three functional parts: First, a sequence, complementary to the conserved regions to which they should match. Second, a barcode, which is unique to each sample. Third, additional nucleotides, which serve as binding sites to the Illumina MiSeq flow-cell. The barcoding of each sample allows simultaneous sequencing of many samples (multiplexing) and the subsequent separation of each sample (de-multiplexing) using computational tools.
16S Microbiome Analysis Applications
- Characterization of bacterial populations in clinical and pre-clinical research
- Taxonomical analysis for research and discovery
- Bacterial species identification for rapid clinical research and infectious disease research
Our 16S Microbiome Analysis Services and Solutions
Sample Preparation - We offer DNA extraction from human, animal and natural environment samples, performed under highly controlled and traceable conditions in our advanced, Copenhagen-based, ISO17025 certified laboratory.
Sequencing – 16S sequencing is performed using the Illumina MiSeq platform with the V3 (2x300bp) reagent kit at highly competitive prices.
Data Analysis - Whether you prefer classic clustering methods (OTUs) or amplicon sequence variants (ASVs with DADA2), our 16S bioinformatics pipelines can be customised to suit your needs:
- Bacterial abundance table using ASVs or OTUs
- Taxonomic profiles (genus/species resolution)
- Biostatistics linking the microbiome to clinical outcomes
- Publication ready figures and statistics
- Integration with study specific datasets
- Customised solutions upon request
Publication ready results – To facilitate your research we deliver your data and results with a easy to read scientific report that summarises the process, the generated data, and the results. To make the report fit into a standard scientific workflow we have structured the report, so it follows the structure of a traditional scientific publication. Also, all raw data is available for you to keep for any potential future use.
At Clinical Microbiomics we have extensive experience managing 16S microbiome analysis projects and offer complete and validated in-house workflows from sample preparation to 16S Amplicon sequencing to data delivery. Your project will be overseen by one of our experienced 16S microbiome study directors who will follow every step of the project workflow, ensuring that quality is consistently at the highest level. You will always be able to contact the study director for direct insights in to the day-to-day processing of your samples.
Timo Kern, PhD, Director of Scientific Operations
If you would like to learn more about our microbiome analysis services using 16S rDNA amplicon sequencing or discuss your study with a member of our science team, please get in touch.