16S/18S/ITS gene sequencing methods are 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). These genes have a high degree of variation even between closely related species.
Conveniently, the these genes consists of both conserved and hyper-variable regions. Universal PCR primers can be designed to target the conserved regions of the genes making it possible to amplify the gene in a wide range of different microorganisms from a single sample. While the conserved region makes universal amplification possible, sequencing the variable regions allows discrimination between specific different microorganisms such as bacteria (16S), fungi (18S, ITS), and archaea (ITS).
- 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/18S/ITS sequencing of microbiota 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.