16S rRNA gene sequencing is commonly used for identification, classification and quantitation of microbes within complex biological mixture such as environmental samples (ex marine water1) and gut samples (ex human gut microbiome2). The 16S rRNA gene is a highly conserved component of the transcriptional machinery of all DNA-based life forms3 and thus is highly suited as a target gene for sequencing DNA in samples containing up to thousands of different species.
Universal PCR primers can be designed to target the conserved regions of 16S making it possible to amplify the gene in a wide range of different microorganisms from a single sample. Conveniently, the 16S rRNA gene consists of both conserved and variable regions. While the conserved region makes universal amplification possible, sequencing the variable regions allows discrimination between specific different microorganisms such as bacteria, archaea and microbial eukarya.
- Environmental Samples (ex soil, water)
- Gut Microbiome
- Sterility Monitoring / Contamination Investigation
- Food Quality Assurance
- Clinical Samples
- Identification efficiency: compared to traditional identification methods such as cloning and/or culturing, 16S rDNA sequencing of bacteria is a faster and more accurate method.
- Dual-zone detection: a comprehensive upgrade to dual-zone (V3 + V4) testing, enables longer sequence and more accurate analysis of colonies.
- Lower cost: much lower requirement of sequencing depth compared to metagenomic sequencing.
- Higher sensitivity: extremely low abundance bacteria can be identified with this method.
On Demand Webinar Viewing
This presentation provides an overview of sample preservation and high quality DNA isolation methods from environmental samples such as soil and fecal samples as well as practical information for performing 16S rRNA sequencing experiments and data analysis. Diverse case study examples will be provided to illustrate the usefulness of this latest application of next-gen sequencing technology to microbiome analysis and environmental science.
- Schmidt TM, Delong EF, Pace, NR (1991). Analysis of a marine picoplankton community by 16S rRNA gene cloning and sequencing. Journal of Bacteriology 173 (14), 4371–4378.
- Ley RE, Bäckhed F, Turnbaugh P, Lozupone CA, Knight RD, Gordon JI. (2005) Obesity alters gut microbial ecology. Proc Natl Acad Sci USA. 102(31), 11070-5.
- Cox MJ, Cookson WO, Moffatt MF. (2013) Sequencing the human microbiome in health and disease. Hum Mol Genet 22(R1), R88-94.