Sequencing and the American Gut Project | Illumina MiSeq, 16S rRNA

Learn about the importance of studying human microbiota and how Next Generation Sequencing (NGS) enables metagenomics research. The American Gut Project is introduced as a citizen science initiative using Illumina technology to survey bacterial communities across thousands of participants.

Participants send biological samples (stool, nasal swab, oral swab, or pet samples) to the lab. DNA is extracted from these samples for downstream analysis.

Learn how 16S ribosomal RNA gene sequencing identifies and characterizes multiple bacterial species simultaneously from complex samples. This method uses conserved and variable regions of the 16S gene to distinguish bacterial taxa without requiring pure culture isolation.

Order amplicon primers for the region of interest and generate amplicon libraries using standard PCR techniques. Add Illumina sequencing and index adapters to prepare samples for sequencing.

Load prepared amplicon libraries onto the Illumina MiSeq benchtop sequencer to generate high-quality sequencing data from all samples in parallel.

Process sequencing data using MiSeq Reporter software's 16S metagenomics analysis pipeline or export to BaseSpace cloud computing environment or third-party software. Generate taxonomic profiles and comparisons of bacterial communities.

Use results to build a knowledge base correlating lifestyle, diet, disease, and bacterial communities. The rapid evolution of sequencing technology makes large-scale microbiome studies feasible and affordable for citizen science projects like the American Gut Project.