Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Modern Molecular Taxonomy01:29

Modern Molecular Taxonomy

527
Advancements in molecular biology have revolutionized the identification and characterization of bacteria, with multiple methods leveraging DNA sequencing for enhanced precision. As sequencing technologies improve and costs decline, these approaches are increasingly used in clinical, environmental, and evolutionary studies.Multilocus Sequence Typing (MLST) examines several housekeeping genes, essential chromosomal genes encoding cellular functions, to distinguish strains. Approximately...
527
Applications of Molecular Taxonomy01:20

Applications of Molecular Taxonomy

456
Molecular taxonomy has revolutionized the understanding and classification of bacteria, providing precise insights into their diversity, evolutionary relationships, and ecological roles. By utilizing molecular techniques such as DNA sequencing and fingerprinting, researchers have made significant strides in various fields related to bacterial studies.Resolving Taxonomic AmbiguitiesMolecular taxonomy has been instrumental in distinguishing closely related bacterial species initially thought to...
456

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Whole-body irradiation causes long-term impairments in the maintenance and function of naïve CD4 T cells specific for self- and non-self-antigens.

Journal of immunology (Baltimore, Md. : 1950)·2026
Same author

IL-17A deficiency in HLA-DR3 transgenic mice enriches beneficial Prevotella species in gut to promote Tregs and reduce CNS autoimmunity.

Microbiome·2026
Same author

Roux-en-Y gastric bypass alters microbial circadian rhythms with links to metabolic improvement.

American journal of physiology. Gastrointestinal and liver physiology·2026
Same author

Age and sex dependent shift in murine gut microbiome reveals pathological links to host cardiovascular and metabolic pathways.

GeroScience·2026
Same author

Public Support for Tobacco Price Promotion Bans Among U.S. Adults: An Online Experiment.

American journal of preventive medicine·2026
Same author

Potential Behavioral Responses After a Hypothetical Ban on Tobacco Price Promotions Among U.S. Adults Who Use Commercial Tobacco.

AJPM focus·2026

Related Experiment Video

Updated: Jan 4, 2026

Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
11:22

Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing

Published on: October 15, 2019

31.0K

Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing.

Shailesh K Shahi1, Kasra Zarei2, Natalya V Guseva1

  • 1Department of Pathology, University of Iowa.

Journal of Visualized Experiments : Jove
|November 5, 2019
PubMed
Summary

This study presents a simplified, high-resolution method for analyzing gut bacteria using 16S rRNA sequencing and DADA2. This workflow aids researchers in microbiome profiling and understanding its role in diseases like multiple sclerosis.

More Related Videos

Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons
10:24

Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons

Published on: August 29, 2014

84.4K
Tick Microbiome Characterization by Next-Generation 16S rRNA Amplicon Sequencing
07:21

Tick Microbiome Characterization by Next-Generation 16S rRNA Amplicon Sequencing

Published on: August 25, 2018

13.3K

Related Experiment Videos

Last Updated: Jan 4, 2026

Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
11:22

Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing

Published on: October 15, 2019

31.0K
Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons
10:24

Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons

Published on: August 29, 2014

84.4K
Tick Microbiome Characterization by Next-Generation 16S rRNA Amplicon Sequencing
07:21

Tick Microbiome Characterization by Next-Generation 16S rRNA Amplicon Sequencing

Published on: August 25, 2018

13.3K

Area of Science:

  • Microbiology
  • Bioinformatics
  • Immunology

Background:

  • The human gut microbiome plays a crucial role in physiological functions and is implicated in inflammatory diseases like multiple sclerosis (MS).
  • Identifying microbial communities associated with disease phenotypes is essential for understanding the microbiome's role in health and disease.
  • Current 16S rRNA sequencing methods for microbiome profiling lack uniform protocols and suffer from low taxonomic resolution, particularly due to short sequencing reads.

Purpose of the Study:

  • To describe a simplified, high-resolution workflow for bacterial profiling of the gut microbiome.
  • To provide a comprehensive guide for researchers using 16S rRNA gene sequencing and established bioinformatics tools.

Main Methods:

  • Two-step amplification of the V3-V4 region of the 16S rRNA gene.
  • Bioinformatic analysis using the Divisive Amplicon Denoising Algorithm-2 (DADA2) pipeline for high-resolution taxonomic assignment (genus and species levels).
  • Utilized freely available analysis tools including DADA2, Phyloseq, and METAGENassist.

Main Results:

  • The described workflow enables high-resolution bacterial profiling, overcoming limitations of previous methods.
  • DADA2 facilitates accurate taxonomic assignment at genus and species levels.
  • The integrated workflow provides a complete solution for microbiome profiling studies.

Conclusions:

  • This simplified and complete workflow offers an excellent tool for researchers conducting microbiome profiling.
  • The method enhances the ability to characterize bacterial diversity in biospecimens.
  • Advancements in sequencing technology and bioinformatics pipelines like DADA2 are crucial for high-resolution microbiome analysis.