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Full-Length 16S/18S/ITS Sequencing


More than 99% of natural microorganisms cannot be isolated and cultured in traditional isolation. Directly from environmental samples, from advanced natural gas technology as a way to explore microbial diversity. The 16S rDNA, 18S rDNA, and internal transcribed spacers (ITS) have been widely used for microbial taxonomy and molecular systematics through a combination of PCR and sequencing methods. We provide full-length 16S/18S/ITS sequencing for microbial diversity analysis of clinical or environmental samples. This method has the ability to discriminate microbial strains even between closely related organisms.

Our Advantages:
  • Long reads (theoretically, the average length of reads can reach 20 Kb, and the maximum reading length can reach Mb level).
  • Lower cost and faster turnaround time.
  • No GC preference caused by PCR reaction; uniform genome coverage.
  • Directly detect base modifications such as DNA methylations.
  • Qualitative and quantitative analysis of full-length transcripts.
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Introduction to Full-Length 16S/18S/ITS Sequencing

The 16S/18S/ITS amplicon sequencing is a precise, sensitive, and cost-efficient tool for microbial diversity analysis and is utilized in thousands of published studies each year. However, this tool is typically restricted to 100-500 bp, which may limit researchers to fragments of preferred taxonomic barcodes. PacBio and Oxford Nanopore have developed novel sequencing platforms that can generate long reads of tens of thousands of nucleotides, which easily span the full length of 16S/18S/ITS. We provide comprehensive full-length 16S/18S/ITS sequencing service based on PacBio SMRT sequencing or Oxford nanopore sequencing platforms for microbial diversity analysis, species classification, and functional profiling.

Full-length 16S/18S/ITS sequencing based on long-read sequencing, as a tool of metagenomics, can achieve species and even strain-level taxonomic resolution, which is difficult for 16S/18S/ITS sequencing based on next-generation sequencing (NGS). Full-length 16S/18S/ITS can provide more information on microbial diversity, phylogeny, and biological function. Full-length 16S/18S/ITS sequencing has been a sufficient value proposition in applications such as characterizing histocompatibility and tracking diversification of HIV env gene.

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

Bioinformatics Analysis

Our bioinformatics analyses include data pre-treatment, taxonomic assignment, diversity analysis, and evolutionary analysis, which are flexible to your specific needs. Please feel free to contact us to discuss your project.

Bioinformatics Pipelines Details
Data Pre-treatment Quality control of the sequences and base calling.
Taxonomic Assignment Operation taxonomic unit (OTU) calling and clustering, rarefaction curve.
Diversity Analysis Shannon index curve, rank abundance curve, heatmap, VENN, PCA, RDA/CCA.
Evolutionary Analysis Construction of phylogenetic trees, estimation of genetic distance.
Custom Analysis Network analysis, correlation analysis, functional analysis, etc.

Sample requirements

Reference

  1. Callahan B J, Wong J, Heiner C, et al. High-throughput amplicon sequencing of the full-length 16S rRNA gene with single-nucleotide resolution. BioRxiv, 2019: 392332.

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