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Microbial diversity analysis
For the analysis of the microbial diversity of complex samples the V3/V4 region of the 16S rRNA (approx. 440 bp) will be amplified [1] and sequenced on an Illumina MiSeq System. A minimum throughput of 200,000 raw reads per sample is guaranteed.
Sequence data will be processed using the QIIME2 software [2] and compared against the SILVA 16S rRNA database [3]. Sequence variants as well as taxonomic identification down to genus level will be reported using QIIME2 visualizations as well as interactive Krona charts [4].
Required material: 1 µg of DNA per sample, minimum 10 samples
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References
- Flocco CG, Methner A, Burkart F, Geppert A and Overmann J (2023) Touching the (almost) untouchable: a minimally invasive workflow for microbiological and biomolecular analyses of cultural heritage objects. Front. Microbiol. 14:1197837. doi: 10.3389/fmicb.2023.1197837
- Bolyen, E., Rideout, J.R., Dillon, M.R. et al. Reproducible, interactive, scalable and extensible microbiome data science using QIIME 2. Nat Biotechnol 37, 852–857 (2019). https://doi.org/10.1038/s41587-019-0209-9
- Christian Quast, Elmar Pruesse, Pelin Yilmaz, Jan Gerken, Timmy Schweer, Pablo Yarza, Jörg Peplies, Frank Oliver Glöckner, The SILVA ribosomal RNA gene database project: improved data processing and web-based tools, Nucleic Acids Research, Volume 41, Issue D1, 1 January 2013, Pages D590–D596, https://doi.org/10.1093/nar/gks1219
- Ondov, B.D., Bergman, N.H. & Phillippy, A.M. Interactive metagenomic visualization in a Web browser. BMC Bioinformatics 12, 385 (2011). https://doi.org/10.1186/1471-2105-12-385