16S rRNA Sequencing for Gut Microbiota Research
Background Services FAQs
Creative Biolabs is one of the largest live biotherapeutic products (LBP) development services suppliers. We have expertise and capabilities in providing high-quality gut microbiota research solutions, including 16S rRNA sequencing service for gut microbiota.
16S rRNA Sequencing for Gut Microbiota
The human gut microbiome is known to influence the health of individuals and is closely related to the occurrence and recovery from diseases. Gut microbiota offers beneficial health effects and, conversely, the compositional disruption of gut microbiota observed in dysbiosis is correlated to various human diseases, including obesity, inflammatory bowel disease, type 2 diabetes, CNS-related disorders, and cancer. In the gastrointestinal microbiome specifically, 16S rRNA profiling has also been widely applied. Many studies have employed 16S rRNA gene sequencing to profile the gut microbiota composition. The most important step in the study of the human gut microbiome is bacterial community profiling; the genomic region encoding the RNA for the ribosomal small subunit (SSU), known as 16S, is the most commonly analyzed. The 16S subunit ribosomal gene is highly conserved between bacteria and archaea and is therefore widely used as a marker gene for assessing microbial phylogeny. The 16S rRNA gene contains nine hypervariable regions (V1-V9) with bacterial species-specific variation, flanked by conserved regions. Thus, amplification in the hypervariable region of 16S rRNA can be used to detect microbial communities in samples, often down to the genus level, or for species-level assignment, if a full-length 16S sequence is retrieved. Due to its high resolution and cost-effective approach, 16S rRNA gene sequencing has become the commonest approach for microbial community profiling of the human gut.
16S rRNA Sequencing Service
Workflows
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Sample Collection and Storage
An important but rarely emphasized aspect of 16S rRNA gene sequencing research is sample integrity. For intestinal microbiota analysis studies, biopsy, surgical tissues, and stool samples are common biological samples collected for characterization. Sample contamination minimization ensures that each biological sample retains as much of its original microbiome as possible and contributes to accurate results of gut microbiome analysis. Short-term and long-term storage conditions after sample transportation are very important to minimize microbiome spectrum differences caused by storage conditions. Low-temperature transportation, such as on ice or dry ice, ethanol-stored, and long-term storage in -80℃ are strongly recommended to maximize microbiome recovery within the sample before nucleic acid extraction.
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Nucleic Acid Extraction
Sample nucleic acid extraction is a simple but critical step in microbial research. Most separation protocols involve three basic steps, including cell lysis, elimination of non-DNA macromolecules, and DNA separation and collection. Gram-positive organisms require stronger lysis conditions due to their thicker cell walls, unlike Gram-negative organisms which require only gentle lysis.
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Sequencing (Selection of Universal 16S rRNA Gene Primers, Sequencing Technologies)
The 16S rRNA gene consists of nine hypervariable conserved regions (V1 to V9) separated by ten highly conserved regions. Most studies were done via sequencing of either one of the V3 or V4 regions, otherwise, a combination of two or more 16S rRNA gene hypervariable regions, whereby the most commonly used were the V3/V4 regions.
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Bioinformatics Analysis
It is important to select the appropriate computing tool for the analysis of the 16S rRNA gene sequencing dataset.
Sequencing Types
Bacterial 16S amplified region
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V4
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V3-V4
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V4-V5
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V5-V7
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Full length
Sample Requirements
Total DNA, ≥300 ng, ≥20 μL, ≥10ng/μL.
Deliverables
Upon completion of the sequencing services, a final report will be issued.
In the rapidly evolving world of LBP, Creative Biolabs' comprehensive portfolio of research and services allows us to help our clients research, discover, and develop new LBP. The human gut microbiota holds promises as a reservoir of novel potential therapeutic targets as well as novel therapeutic agents, paving a new and exciting avenue in drug discovery. If you are interested in our 16S rRNA sequencing services for gut microbiota research, please contact us for more details.
Frequently Asked Questions
What are the research applications of 16S rRNA gene sequencing?
16S rRNA gene sequencing is extensively used in research due to its ability to provide detailed insights into microbial communities. Here are some key research applications:
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Microbial Diversity Studies: Researchers use 16S rRNA sequencing to explore the diversity of microbial communities in various environments, such as oceans, soils, and the human gut.
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Microbiome Analysis: It helps in studying the composition and function of microbiomes in different hosts, including humans, animals, and plants, which is crucial for understanding health and disease.
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Ecological Studies: This technique is used to investigate the roles of microbes in ecosystems, including nutrient cycling, decomposition, and symbiotic relationships.
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Evolutionary Biology: Researchers use 16S rRNA sequencing to study the evolutionary relationships among different microbial species, helping to construct phylogenetic trees.
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Biotechnology and Industrial Research: It aids in identifying and characterizing microbes involved in industrial processes, such as fermentation, bioremediation, and bioenergy production.
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Pathogen Identification: In research settings, it is used to identify and study pathogenic bacteria, contributing to the development of new diagnostic methods and treatments.
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Antibiotic Resistance Studies: Researchers use it to monitor the spread of antibiotic-resistance genes within microbial communities, which is vital for public health.
What advantages of 16S rRNA gene sequencing?
16S rRNA gene sequencing offers several advantages, making it a popular choice for studying microbial communities. Here are some key benefits:
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Broad Range of Detection: It can identify a wide variety of bacteria and archaea, including those that are difficult or impossible to culture in the lab.
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High Resolution: Provides detailed information about the genetic makeup of microbial communities, allowing for precise identification and classification.
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Cost-Effective: Compared to whole-genome sequencing, 16S rRNA sequencing is more affordable, making it accessible for large-scale studies.
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Rapid and Efficient: The process is relatively quick and can be automated, enabling high-throughput analysis of multiple samples simultaneously.
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Phylogenetic Insights: Helps in constructing phylogenetic trees, providing insights into the evolutionary relationships among different microbial species.
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Non-Culture Based: Does not require culturing of microbes, which is particularly useful for studying environments where many organisms are not easily cultured.
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Versatile Applications: Can be used in various fields, including clinical diagnostics, environmental microbiology, industrial processes, and more.
For Research Use Only. Not intended for use in food manufacturing or medical procedures (diagnostics or therapeutics). Do Not Use in Humans.
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