Resolve the full 16S rRNA gene with PacBio HiFi Sequencing and our bioinformatic analysis.
Standardized procedures with rigorous QC with Microbial Standards provide accurate and reliable data.
Receive Full-Length 16S sequencing results in as little as 3 weeks
The Full-Length 16S rRNA sequencing service report provides compositional bar plots, composition heatmaps, alpha-diversity and beta diversity plots, and statistical group comparison with LEfSe. Our team of expert bioinformaticians are here to help you get the most out of your data.
View Sample ReportZymoBIOMICS Microbial Community DNA Standard (D6305) was used as a benchmark control. Zymo Research Full-Length 16S Sequencing provides the same bacterial profiling as defined in our Microbial Community DNA Standard.
Zymo Research’s experienced team of scientists are here to answer your questions and work with you to create “end-to-end microbiome sequencing & analysis” for your goals. Obtain consultation for your project with the latest biotechnology today.
| Sequencing Platform | PacBio Sequel Ile |
|---|---|
| Sequencing Kit | SMRT Cell 8M |
| Target Amplicon | The full-length 16S rRNA gene is amplified with 27f (AGRGTTYGATYMTGGCTCAG) and 1492r (RGYTACCTTGTTACGACTT) |
| Accepted Sample Types | Extracted genomic DNA, or raw samples |
| DNA Extraction |
ZymoBIOMICS 96 Magbead DNA (D4308) ZymoBIOMICS DNA Miniprep (D4300) |
| Library Prep | We follow PacBio recommended protocol (https://www.pacb.com/wp-content/uploads/Procedure-checklist-Amplification-of-bacterial-full-length-16S-rRNA-gene-with-barcoded-primers.pdf) |
| 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 is used 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 Visualization | 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 |
Targeted sequencing amplifying certain variable regions of 16S rRNA genes has become a popular and routine technique for microbiome composition profiling. Traditionally, researchers relied on short-read sequencing platforms (e.g. Illumina MiSeq) for this application. However, short-read sequencing read-length restricts this application to one or a few variable regions of 16S rRNA genes, e.g. V4, V3-V4, V1-V3, V1-V2 and V4-V5. As a result, the taxonomy resolution is limited.
With the introduction of long-read sequencing platforms (e.g PacBio Sequel II and Oxford nanopore sequencing), researchers can now leverage the complete taxonomy resolution power of the full 16S rRNA gene. Full-Length 16S Sequencing is the latest in this long-read technology.
Zymo Research is a frontier in microbiome analysis and is one of the few Full-Length 16S Sequencing service providers. In light of our motto to “Make Science Simple”, we are happy to be on the forefront of Full-Length 16S Sequencing customer offerings. Zymo Research is happy to combine PacBio HiFi sequencing and error correction bioinformatics analysis (e.g. Dada2), so that we can recover the error-free full-length 16S sequences from a mixed microbial community (e.g soil or feces). We are offering this powerful Full-Length 16S Sequencing service for microbiome researchers and industry partners alike. As a Certified PacBio Provider you can rest assured that your samples will be well taken care of as they undergo Full-Length 16S Sequencing.
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