Microbiome analysis is performed via 16S rRNA gene sequencing. 16S sequencing is commonly used for identification, classification and quantitation of microbes within complex biological mixture such as human gut microbiome samples1). The 16S rRNA gene is a highly conserved component of the transcriptional machinery of all DNA-based life forms2 and thus is highly suited as a target gene for sequencing DNA in samples containing up to thousands of different species.
How It Works:
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.
Other Applications of 16S Sequencing:
- Environmental Samples (ex soil, water)
- Sterility Monitoring / Contamination Investigation
- Food Quality Assurance
Technical Advantages of 16S Sequencing
- 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.