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.
Why use 16S profiling?
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
1. Initial meeting
2. Sample delivery
- Validated input: ≥ 10pg / sample
- Minimum volume: 15μL / sample
- Quality: Column or bead purified DNA
3. Sample entry QC
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
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.
- TAT: 2-3 weeks after successful sample QC
- FastQ files via secure e-transfer
- Quality score Q30 of ≥80% for PE150 reads
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.
We have summarized key information about our 16S microbial profiling service into a service specification sheet.
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