Data processing and microbiome profiling

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All raw sequencing data must be processed before microbiome profiles can be generated and biologically relevant information can be obtained. This includes evaluation of the data quality and filtering of low quality data. After performing quality filtering of the sequencing data, it is time to generate the microbiome profiles. A taxonomic microbiome profile contains information on which taxa are found in the community and their relative abundances*.

If shotgun metagenomic sequencing is used (and not marker gene based sequencing), it is also possible to profile the functional capacity of the microbiome community. This provide information on the individual genes found in the community and an estimate of the functional pathways they contribute to.

*If a spike-in solution is included in the sequencing, absolute abundance taxonomic profiles will be generated and analyzed alongside the relative abundance data.

What we do

Data processing and quality evaluation

  • Evaluate the quality controls where applicable
  • Evaluate the quality of the raw sequencing data
  • Filter the data based on the quality evaluation. Quality filtering include removal of (i) adaptors, (ii) low quality data and (iii) contaminants and host reads (where relevant)

Microbiome profiling

  • We take quality filtered data and generate microbiome profiles. The specific software and reference data we use depend on the type of data. For example, different software solutions are used to profile 16S and shotgun metagenomic data, and different software and/or reference data will be used to profile stool, saliva or soil samples. The decisions we make also in part depend on the goal of the project, such as the selection of marker-based or assembly-based approaches for shotgun data.

What you get

For each project, a private project folder is made available on BIOMCARE server, where all resulting data, statistical results, illustrations, reports etc. are available for download.

Data

  • Quality filtered data in fastq format

Microbiome profiles

  • Taxonomic profiles for amplicon data
  • Taxonomic and functional profiles for shotgun data

Diversity measures

  • Alpha diversity (including observed and extrapolated species/genera richness).
  • Beta diversity

Illustrations

  • Illustrations that support data evaluation and further analysis decisions. The illustrations include ordination plots and stacked barplots.

Reports

1) Data Quality Report, including:

  • A description of the data quality
  • Details of the performed quality filtering.
  • Illustrations that support easy overview of reads/base pairs removed at different steps and number reads remaining post filtering.

2) Microbiome Profile Report, including:

  • Details of the microbiome profiling performed
  • Explanations to the provided tables and illustrations
  • Summary of our evaluations and considerations (e.g. do we see potential issues, how we suggest samples and taxa should be filtered in further analysis)

Personal guidance and communication

A key factor for successful outcomes of every project is communication. Every project is different, and good personal communication allow us to understand exactly what our customers wish to achieve. This is especially essential when we design the statistical analysis and incorporate customer provided information. We typically have the most active communication at the beginning where samples are being collected and shipped, and then again when statistical analysis are initiated, and results starts to emerge. At the end of the project, we present the project results in an online or face-to-face meeting, depending on geographical circumstances and project size/complexity.

The communication was swift and the samples rapidly analysed.

Sidsel Støy

M.D., Hepatology and Gastroenterology, Aarhus University Hospital