From extraction to sequencing results in as little as 2 weeks
Species-level resolution with curated 16S rRNA database
Get more valuable insights from your sample
All projects include a comprehensive report with bioinformatics analysis and raw sequencing data. The 16S rRNA sequencing service report 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. Our team of expert bioinformaticians are here to help you get the most out of your data.
| Species | Zymo Research 16S Pipeline | Other Service Providers |
|---|---|---|
| Listeria monocytogenes | ||
| Staphylococcus aureus | ||
| Enterococcus faecalis | ||
| Lactobacillus fermentum | ||
| Escherichia coli | ||
| Salmonella enterica | ||
| Pseudomonas aeruginosa | ||
| Bacillus subtilis | ||
| Species-level Resolution |
By combining leading edge sequencing error correction methods with a highly curated 16S rRNA database, our 16S amplicon sequencing service can provide taxonomic identification down to the species level.
Relative abundance alone cannot define changes in a microbial community. In cases like comparing treatment or therapeutic effects, it can be just as important to know how cell numbers are changing. With absolute abundance quantification provided by our 16S amplicon sequencing service, open new insights into the microbial community dynamics of your project.
Our 16S amplicon sequencing service is continually monitored with mock community controls to ensure data accuracy and consistency. Don’t let run-to-run variations interfere with your long-term study or project.
| Sequencing Platform | Illuminia MiSeq ™, NextSeq2000 ™ |
|---|---|
| Read Length | 2 x 300 bp, Unique Dual Indexing (UDI) |
| Target Regions |
V1-V2, V1-V3, V3-V4, V3, V4, V4-V5, V5-V6, V6-V8, ITS2, 18S Others available upon 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 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 |
Yes. The sample requirements and submission guidelines are the same as for our 16S Sequencing Service.
We have processed a wide variety of sample types, from human tissue samples, environmental samples, to food samples. If you have questions or concerns, please feel free to fill out an inquiry or contact us.
We recommend sending 200-500 mg for solid samples or 500-1mL of liquid samples in general. This may vary depending on the specific sample type.
The minimum volume of DNA required is 30 µl with a recommended concentration of 10-50 ng/µl. If the concentration is lower, we can often still successfully sequence as our pipeline is quite robust.
There is no sample minimum required in order to submit to our 16S Sequencing Service.
Because of our included quality control standard, it is possible to batch samples over time and have confidence that the samples were processed in the same way. We recently completed a project that had a total of >2,000 samples processed over a period of 6 months. By comparing the results from the standard for each group of samples, we had confidence that the results from different batches could be compared together.
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