Amplicon sequencing is a next-generation sequencing method used to identify bacteria, archaea and fungi from any microbiome sample. The method uses primers designed to target conserved regions of ribosomal RNA and amplify the the hypervariable regions between them for sequencing. These hypervariable regions are unique enough to resolve the community composition down to the genus, and sometimes species, level.
Zymo’s sequencing services begin with a free consultation and their workflows can be fully customised to your project’s needs. The team is highly skilled and experienced in producing high-quality data from your sequencing samples and providing customised, user-friendly bioinformatics analysis reports. For a limited time Zymo are offering your first ten 16S analyses free!
What you can expect from Zymo’s services
- Industry-leading turnaround
- Unrivalled data quality
- Stringent QC using microbial standards
- No minimum sample number required
- Absolute abundance quantification
- Experience across a huge range of sample types
- User-friendly analysis report
Comprehensive bioinformatics report
Zymo’s analysis report includes absolute abundance quantification, giving you insight into changes in the number of cells within the community you are analysing and making it easier to draw conclusions in cases of comparing treatment or therapeutic effects.
Zymo’s amplicon sequencing service report also provides compositional bar plots, taxonomy heatmaps, alpha-diversity and beta diversity analyses. Group comparisons and LEfSe biomarker discovery are also reflected in all 16S/ITS amplicon sequencing service reports. Zymo’s team of experts are on hand to help you get the most out of your sequencing data.
Continuous monitoring for consistency and standardisation
Zymo’s amplicon services are continuously monitored with their own microbial community standards for batch-to-batch consistency. The standards are the some of the most highly cited in the industry and have helped the microbiomics research community ensure reproducibility and data quality in over 1000 publications.
16S/ITS amplicon sequencing technical sepcifications
| Sequencing platform | Illumina MiSeq ™, NextSeq2000 ™ |
|---|---|
| Read length | 2x300 bp, unique dual indexing (UDI) |
| Target regions | V1-V2, V1-V3, V3-V4, V3, V4, V4-V5, V5-V6, V6-V8, ITS2, 18S. Other available on request |
| Accepted sample types | Extracted genomic DNA or raw samples |
| DNA extraction |
ZymoBIOMICS 96 MagBead DNA (D4308) ZymoBIOMICS DNA Miniprep (D4300) |
| Library prep |
Quick-16S NGS Library Prep (D6400) Quick-16S Plus NGS Library Prep (D6421) |
| Plastid rRNA blockers | PNA PCR blockers - blocks amplification of the V4 region of chloroplast and mitochondrial rRNA genes |
| Absolute quantification | Quantification of absolute 16S or ITS gene copy number using qPCR with library preparation primers |
| Quality control | A positive control for DNA extraction (ZymoBIOMICS Microbial Community Standard, D6300) and a positive control for library preparation (ZymoBIOMICS Microbial Community DNA Standard, D6305) are included in each project as a quality control. Results are reported in the final project report. Negative extraction controls and negative qPCR controls are also monitored with real time PCR and can be analyzed upon request. |
| Sequencing analysis | Amplicon Sequence Variant (ASV) based analysis with DADA2 and Qiime 1.9 to provide higher accuracy and better resolution of data. |
| Reference database | As publicly available databases (e.g. Greengenes, UNITE) contain a multitude of errors, inconsistent annotations, and poor sequence quality, we utilized an in-house curated reference database. |
| Data visualisation | Absolute abundance boxplot, taxa abundance barplot, taxa abundance heatmap, ASV abundance heatmap, alpha diversity boxplot and rarefaction curves, beta diversity 3D Emperor plot, Taxa2ASV decomposer |
| Statistical analysis | LEfSe analysis uncovers taxa/traits that are differentially distributed among defined sample groups. |
| Data storage | Raw sequencing data and analysis report are stored for up to 3 months |
Material for download
Zymo Research targeted service sample report
Zymo Research shotgun service sample report
Tell us about your project
To discuss a custom project, please contact our Zymo Research specialists
