Full-Length 16S/18S/ITS Amplicon Sequencing

CD Genomics is a leading provider of genomic services. We have extensive experience in long-read amplicon sequencing using the PacBio platform. PacBio SMRT long-read sequencing is capable of generating complete, uniform, unbiased coverage spanning long amplicons, allowing for a more realistic restoration of the microbial community structure in the samples.

Introduction of 16S/18S/ITS Amplicon Sequencing

16S/18S/ITS amplicon sequencing, also known as microbial diversity sequencing, refers to the sequencing of PCR products from specific regions such as 16S ribosomal DNA (rDNA), 18S rDNA, Internal Transcribed Spacer (ITS), and functional genes. This method does not require culturing microorganisms, and by detecting sequence variation and abundance in the target region, information on microbial community structure, evolutionary relationships, and microbial relevance to the environment can be obtained from environmental samples.

16S and 18S rDNA are the most commonly used marker genes in metataxonomics. Among them, 16S rRNA gene (~1,500bp) with nine variable regions interspersed between conserved regions, is used to analyze the diversity of bacteria or archaea. 18S rRNA gene (1,500-2,000bp) is utilized to reflect species differences in eukaryotes in a given sample. While ITS (400-900bp) is the spacer DNA between the small-subunit and large-subunit rRNA genes in bacteria, fungi, and archaea. ITS region has been widely utilized for the classification of fungal communities, including ITS₁ and ITS₂.

Advantages of Full-Length 16S/18S/ITS Amplicon Sequencing Analysis

PacBio SMRT long-read sequencing has overcome the limitations of short reads, allowing researchers to obtain full-length reads of 16s rDNA in bacteria or 18S rDNA and ITS regions in eukaryote.

• High throughput and high-fidelity data: PacBio Sequel's circular-consensus-sequencing (CCS) analysis for generating high accuracy single-molecule consensus sequences of large genomic regions at one time.
• Improved resolution of species identification: High sequence assignment rates for annotation to each taxonomic level.
• Improved accuracy of species identification: Achieving species-level identification.

Sample Requirements

1. Sample Type:gDNA/PCR Products, OD260/280=1.8~2.0, no degradation and no contamination.
2. DNA amount:
   -gDNA ≥100 ng
   -PCR fragment ≥500 ng
3. Sequencing strategy:
   PacBio Sequel platform, 2000~20000 CCS reads/per sample

Workflow of Our Service

Analysis Workflow and Contents

Standard Analysis	Advanced Analysis
-OTU clustering and annotation -Species annotation -Species composition analysis -Alpha diversity analysis -Beta diversity analysis -Functional prediction -Multi statistics	-Species variation analysis -Correlation analysis of environmental factors -Phylogenetic analysis -Customized analysis

If you are interested in our full-length 16S/18S/ITS amplicon sequencing service, please feel free to contact us. We've got everything covered for your needs and are ready to assist.