Related Experiment Video
Updated: May 17, 2026
11:22
Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
Published on: October 15, 2019
Nucleic acid-based methods to assess the composition and function of the bowel microbiota
Blair Lawley1, Gerald W Tannock
1Department of Microbiology and Immunology, University of Otago, Dunedin, New Zealand.
Gastroenterology Clinics of North America
|October 30, 2012
Summary
This article reviews nucleic acid-based methods for analyzing bowel microbiota composition and function. Researchers should use multiple validated techniques for comprehensive analysis, starting with screening to guide in-depth studies.
Area of Science:
- Microbiology
- Molecular Biology
- Genomics
Background:
- The gut microbiota plays a crucial role in human health.
- Understanding microbiota composition and function is essential for disease research.
- Nucleic acid-based methods offer powerful tools for microbiota analysis.
Purpose of the Study:
- To describe various nucleic acid-based methods for assessing bowel microbiota.
- To highlight the accessibility and validation status of different techniques.
- To provide guidance on selecting appropriate methods for microbiota research.
Main Methods:
- Polymerase chain reaction (PCR) for targeted analysis.
- Sequencing-based methods for comprehensive profiling (e.g., 16S rRNA sequencing).
- Metatranscriptomics for functional analysis of gene expression.
Main Results:
- A range of methods exist, from simple to complex.
- Validated tools are available for most researchers.
- No single method is sufficient for complete microbiota analysis.
Conclusions:
- Multiple, validated nucleic acid-based methods are recommended for robust microbiota studies.
- Initial sample screening can optimize resource allocation for in-depth analysis.
- A combination of techniques provides a more complete understanding of the bowel microbiota.
Related Concept Videos
01:19Methods to Assess Microbial Communities
Microbial communities, comprising bacteria, archaea, and eukaryotic microorganisms, inhabit diverse ecosystems and play crucial roles in environmental and biological processes. Their diversity is defined by three main parameters: species richness (the number of distinct species), species abundance (the relative quantity of each species), and species evenness (how uniformly individual species are distributed in various locations). These factors together shape the structure and ecological balance...
01:30Methods to Assess Microbial Populations
Assessing microbial populations is crucial for understanding microbial roles in health, ecology, and industry. Various complementary techniques—both culture-based and molecular—enable detailed analysis of microbial abundance, diversity, and function.Viable Plate CountThe viable plate count is a traditional culture-based method used to estimate the number of living microbes in a sample. After serial dilution, the sample is spread onto nutrient agar plates. Each viable cell forms a visible...
01:29Modern Molecular Taxonomy
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...
01:30Serum Laboratory Studies, Stool Test, Breath Test
Gastrointestinal (GI) diagnostic studies are pivotal in confirming, ruling out, diagnosing, or staging various diseases, including cancers. Following diagnosis, allocating time for discussions with the patient and providing informational resources is crucial. Diagnostic assessments of the GI tract often occur in outpatient settings like endoscopy suites or GI labs. Preparation for these tests may include dietary restrictions, fasting, liquid bowel preparations, laxatives, enemas, and the...
01:18Functions of the Gut Microbiota
The gut microbiota includes trillions of microorganisms that colonize the human gastrointestinal tract, including bacteria, archaea, viruses, and fungi. This complex ecosystem plays a critical role in maintaining intestinal and systemic health. Most of these microbes inhabit the large intestine, establishing a relatively stable and diverse community that contributes to gut homeostasis through various metabolic, immunological, and protective mechanisms.Dominant bacterial phyla, such as...
01:28Methods of Classification and Identification
Bacterial identification relies on a diverse array of techniques to classify and understand microorganisms, each tailored to uncover specific characteristics. Traditional morphological approaches, while still valuable, are limited for closely related or structurally simple organisms. Modern methods integrate biochemical, serological, genetic, and advanced molecular tools to achieve greater accuracy.Morphological and Biochemical TechniquesMorphological characteristics, such as cell shape and...