The 16S rRNA gene is present in all bacteria and archaea and serves as a means of identification based on the individual sequence allowing comparison of taxonomic diversity within a sample or identifying the presence of specific bacteria of interest. Oxford Nanopore Technologies’ 16S kit targets the whole gene, not just the hypervariable regions so there is more taxonomic context with which to assign a classification to the read, improving precision in metagenomic species identification.
Why do 16S sequencing using Oxford Nanopore’s 16S Barcoding kit? Oxford Nanopore devices can sequence long reads, so targeting the whole 16S rRNA gene with a single amplicon is easy and simple.
- Prepare, sequence, analyse: an end to end workflow to assess metagenomic diversity.
- PCR Primers are optimised for sequencing on Oxford Nanopore devices.
- No optimisation of multiple primers to cover the hypervariable regions is required
- Easy amplification of target within a background of host
- Quick workflow –5 minutes post PCR to prep
- Up to 24 individual barcodes available, reducing costs per sample.
- Simple to use analysis workflow to provide a report of what your microbiome contains.
This kit is recommended for users who
- wish to multiplex samples to reduce price per sample
- want to do 16S sequencing
- are interested in genus level bacterial identification
Barcoding or multiplexing is useful when the amount of data required per sample is less than the total amount of data that can be generated from a single flow cell: it allows a user to pool multiple samples and sequence them together making more efficient use of the flow cell.
The 16S Barcoding Kit offers a method of amplifying and barcoding the ~1500 bp 16S rRNA gene from multiple samples and sequencing them together. By narrowing down to a specific region of interest, a user can see all the organisms in the sample without sequencing unnecessary regions of the genome, making the identification quicker and more economical.
The kit is supported by the EPI2ME 16S-BLAST workflow, which can be used to analyse data from the 16S protocol. Deconvolution of barcoded sequencing data is supported by Guppy and EPI2ME which classify the barcode sequence and sort reads into corresponding folders.