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Bacterial 16S rRNA Sequencing

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Bacterial 16S rRNA sequencing utilizes consensus primers to specifically target conserved regions of the bacterial 16s ribosomal RNA (rRNA) gene followed by high-throughput sequencing. We guarantee the quality of our data and report.

Our Advantages:
  • The experts will work closely with you throughout the entire service lifecycle.
  • Expertise in dealing with various types of environmental and human microbiome samples.
  • Accredited lab with the highest standard of proficiency and quality assessment at every step of the process.
  • Fast turnaround time and highly experienced staff.
  • Next-generation sequencing (NGS) and third-generation sequencing platforms.
Tell Us About Your Project

We are dedicated to providing outstanding customer service and being reachable at all times.

Request a Quote

Introduction to our Bacterial 16S rRNA Sequencing

The 16S rRNA gene, a housekeeping gene in almost all bacteria, is about 1550 bp long and consists of both conserved and variable regions. The sequence of the variable region can be used for bacterial taxonomy and phylogeny. 16S rRNA sequencing generally involves the selective PCR amplification using appropriate primers that target the16S rRNA gene, followed by next/third generation sequencing through Illumina MiSeq/HiSeq, Roche 454, or Ion Torrent instruments, or by third-generation sequencing through PacBio SMRT or Nanopore instruments. Coupled with powerful bioinformatics tools, we can perform basic sequence processing, diversity analysis, taxonomic assignment, and community comparisons. Our robust 16S rRNA analysis pipeline for phylogenetic assignment is using three popular databases – Silva, Green Genes, and Ribosomal Database Project (RDP).

Bacterial 16S rRNA sequencing can determine the microbial diversity, microbial abundance, and possible microbial functions. It has played an important role in the identification of the global bacteria communities in both healthy and diseased groups for disease diagnosis, biomarker discovery, and target discovery. Bacterial 16S rRNA sequencing has also been applied to areas such as environmental conservation, agricultural production, petroleum exploration, and industrial manufacturing.

Bacterial 16S rRNA Sequencing Workflow

  1. Consultation: It is essential to have an initial consultation with our experts regarding the samples and the project to define objectives and set clear outcomes.
  2. Sample Submission: We will provide instructions on how to handle, process, and transport the samples and specify sample requirements based on sample type.
  3. DNA Quality Control (QC): We assess the DNA on quantity, quality, and purity prior to library preparation.
  4. Library preparation: We can construct various 16S rRNA libraries that are suitable for different sequencing platforms.
  5. Sequencing: We offer comprehensive 16S rRNA sequencing services by using the next/third generation sequencing technology.
  6. Bioinformatics analysis: Complete bioinformatics analysis services, including raw data processing, diversity analyses, taxonomic assignment, community comparisons, and etc.
  7. Final Report: A comprehensive report is the final deliverable in a timely fashion.

Bioinformatics Analysis

After DNA sequences are obtained, bioinformatics tools such as QIIME and mothur are commonly used for raw data processing, taxonomic assignment, diversity analysis, community comparisons, and evolutionary analysis.

Pipeline The problem to be solved
Basic data processing Filtering and trimming of poor-quality sequence
Taxonomic assignment Operation Taxonomic Unit (OTU) clustering and species annotation
Diversity analysis Classification and abundance analysis of single species
Community comparisons Measure the difference in bacterial community composition among different samples, and conduct network analysis and correlation analysis
Evolutionary analysis Construction of phylogenetic tree

Sample Requirement

    1. DNA sample: DNA > 20 ng (no host, impurities or contamination).
    2. No obvious degradation and protein contamination, 1.8 < OD260/280 < 2.0, OD260/230 ≥1.8, concentration ≥30 ng/μL.

Sampling kits: We provide a range of microbial sampling kits for clients, including MicroCollect™ oral sample microbial collection products and MicroCollect™ stool sample collection products.

Deliverables: Raw sequencing data (FASTQ), trimmed and stitched sequences (FASTA), data QC report, and custom bioinformatics analysis report.

* For Research Use Only. Not for use in diagnostic procedures or other clinical purposes.

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