The amplification of the 16S prokaryote-specific gene allows you to analyze the microbiome together with the host without any loss of efficiency or sequencing depth. GenomeScan’s scientists have designed an optimized library preparation method, leading to a higher yield than other kits available on the market. A dual-indexing strategy ensures that hundreds of samples can be analyzed together, making this a very reliable and powerful tool.
16S microbial profiling
Uncover taxonomic classification of microbes in complex mixtures
Why use 16S profiling?
Discuss your project with me
How 16S profiling works
Identification and comparison of the microbes present in your samples is performed by amplification of the fourth hypervariable domain (V4) of the 16S ribosomal RNA gene. The 16S region is a conserved region of the prokaryotic ribosome but it also contains hypervariable sequences that provide the species-specific signature.
- Taxonomic classification in complex mixtures
- Detailed overview of the prokaryotic composition to family, genus and sometimes species level
- Relative frequencies in comprehensive report
- Publication-ready figures
Service workflow
1. Initial meeting
When you consider using one of our services, our scientific team will help choosing the optimal project design. Each service can be customized by adding options. You will receive an overview of project budget and deliverables, so you can make an informed choice. Once you return your Purchase Order form, your project manager will contact you to discuss the next steps.
2. Sample delivery
Sample specifications for 16S microbial profiling are:
Purified DNA
- Validated input: ≥ 10pg / sample
- Minimum volume: 15μL / sample
- Quality: Column or bead purified DNA
3. Sample entry QC
After sample arrival, an entry QC is performed to determine if the sample quality is sufficient so we can guarantee the data outcome stated in your quotation. You will receive a report with sample QC results to inform you of our findings before proceeding further. Samples that fail QC can be replaced by spare samples delivered in the same shipment or sent at a later stage.
4. Library prep/QC and sequencing
The library resulting from sample preparation goes into a library QC step to ensure sufficient material per sample and the right library size. Sequencing on Illumina NovaSeq 6000 (PE150):
- Standard read depth 200K PE150 reads / sample
- Unique dual indexed sequencing adapters
5. Data QC
The raw data that is generated by the sequencer must be divided to obtain one data file per sample. This is called demultiplexing.
Your project manager checks the data quality by analyzing the quality metrics of the run. Final inspection of the data takes place and the dataset is transferred onto a hard disk.
6. Reporting
You receive your Project Report together with your data. It contains an overview of the experimental methods used, sample specifications, and a summary of all technical information for publication. Deliverables:
- TAT: 2-3 weeks after successful sample QC
- FastQ files via secure e-transfer
- Quality score Q30 of ≥80% for PE150 reads
Data analysis
GenomeScan BioIT experts have set up a data-analysis pipeline to efficiently process all microbiome data. Sequential steps include data trimming and preparation for alignment to the NCBI reference database to determine the characteristics and relative abundancies of different microbial populations.
The output consists of a comprehensive table containing the various microbes that were detected. It displays the taxonomic rank, genome size and number of reads that classifies the various microorganisms. Krona-plots provide an interactive tool to visualize the composition of the sample.
Service specifications
We have summarized key information about our 16S microbial profiling service into a service specification sheet.
Let's get the conversation started for your next NGS project
Please either fill in this form or email us directly at info@genomescan.nl
and we will get in touch with you to discuss your requirements.
