Full-length 16S Amplicon Sequencing

The sequence contains several conserved and hypervariable regions. Among them, the genus or species-specific hypervariable regions are the most suitable indicator for bacterial phylogenic and taxonomic identification. Combining the long-read advantages of the third-generation sequencing platform, the full-length 16S sequence can be sequenced, and species annotation information at the level of Species and even strains can be obtained, which greatly improves the resolution of data analysis, so it has gradually become a research in the field of microorganisms. One of the important strategies for environmental microbial diversity and community composition differences.
The raw data from the sequencing machine will be used with PacBio standard software to correct Circular consensus sequence (CCS) and generate CSS reads.
The corrected CCS sequence is used as the beginning of the analysis, followed by removing the primers (Primer) and performing DADA2 denoising analysis. After denoising analysis, ASVs (Amplicon Sequence Variants) are obtained, followed by species classification analysis, including species abundance, Alpha diversity calculation, Beta diversity evaluation, Venn diagram, etc., to obtain species richness and evenness information in the sample, common and unique ASVs information among different samples or groups.
  1. Amount:

    1. Genomic DNA ≧ 500 ng

    2. Concentration ≧ 30 ng/ ul (Qubit®)

    3. Volume ≧ 20 ul

  2. Purity:
    OD260/OD280 = 1.8-2.0
    No degradation and contammination
PacBio Sequel IIe

Courtesy of PacBio of California, Inc.