What Are 16S and ITS rRNA Sequencing?
16S and Internal Transcribed Spacer [ITS] ribosomal RNA [rRNA] sequencing are common amplicon sequencing methods used to identify and compare bacteria or fungi present within a given sample. Next-generation sequencing [NGS]-based ITS and 16S rRNA gene sequencing are well-established methods for comparing sample phylogeny and taxonomy from complex microbiomes or environments that are difficult or impossible to study.
The prokaryotic 16S rRNA gene is approximately 1500 bp long, with nine variable regions interspersed between conserved regions. Variable regions of the 16S rRNA gene are frequently used for phylogenetic classification of genus or species in diverse microbial populations.1 The ITS1 region of the rRNA cistron is a commonly used DNA marker for identifying fungal species in metagenomic samples.2
16S Sequencing Method Guide
Amplicon-based next-generation sequencing of the 16S gene offers several advantages over capillary sequencing or PCR-based approaches. Learn how it works with this guide to 16S sequencing methods.
Advantages of NGS-Based ITS and 16S rRNA Analysis
A key benefit of 16S and ITS ribosomal RNA NGS methods is that they provide a cost-effective technique to identify strains that may not be found using traditional methods. Unlike capillary sequencing or PCR-based approaches, next-generation sequencing is a culture-free method that enables analysis of the entire microbial community within a sample.
16S rRNA NGS allows microbiologists to achieve genus-level sensitivity for metagenomic surveys of bacterial populations. ITS analysis with NGS enables rapid fungal identification to help advance our understanding of the mycobiome. Furthermore, NGS offers the ability to combine multiple samples in a sequencing run.
Scientists Discuss 16S rRNA Sequencing
The Saca la Lengua project used 16S and 18S rRNA sequencing to identify the bacteria and fungi that live in the human mouth.
Read InterviewMichael Bunce, PhD uses next-generation sequencing and metabarcoding methods to study environmental DNA [eDNA].
Read InterviewPhil Hugenholtz, PhD explains the difference between 16S rRNA and shotgun metagenomic sequencing, and describes how NGS has made a difference in his research.
Read Interview
Applications of 16S and ITS rRNA Sequencing
Using the 16S metagenomics workflow with the iSeq 100 System, you can achieve genus-level sensitivity for surveys of bacterial populations.
Read Application Note
Get genus-level detection of fungi in diverse sample types with the ITS metagenomics workflow.
Read Application Note
Demonstrated 16S and ITS rRNA NGS Protocols
Workflow for ITS and 16S rRNA Sequencing
Illumina offers products to support NGS-based 16S and ITS rRNA analysis studies, from library preparation to data analysis and interpretation. Our user-friendly workflow can help take the guesswork out of your experiments.
Click on the below to view products for each workflow step.
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The iSeq 100 System makes next-generation sequencing easier and more affordable than ever.
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The MiSeq benchtop sequencer enables targeted and microbial genome applications, with high-quality sequencing, simple data analysis, and cloud storage.
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Prepare sequencing libraries for small genomes, amplicons, plasmids, and other applications.
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Illumina NGS is enabling "citizen science" metagenomics studies of the human microbiome.
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A detailed look at our demonstrated 16S rRNA analysis workflow, including data visualization and success stories.
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