The 16S Barcoding Kit delivers rapid, genus-level bacteria identification using nanopore sequencing.

• Sequence and analyse the complete 16S rRNA gene for improved taxonomic resolution
• Multiplex samples to reduce price per sample
• Analyse samples using the EPI2ME 16S workflow or your own analysis pipeline

The Oxford Nanopore 16S Barcoding Kit offers a method of amplifying and barcoding the entire ~1500 bp 16S rRNA gene extracted gDNA, followed by library preparation. 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. Analysing the full 16s rRNA gene has been shown to be more informative than analysing a subset of exons.

The DNA is amplified by PCR using specific 16S primers (27F and 1492R) that contain 5’ tags which facilitate the ligase-free attachment of Rapid Sequencing Adapters.

The 16S Barcoding Kit 24 V14 has been upgraded to use Kit 14 chemistry, which includes improved median raw read sequencing accuracies with high output on the latest nanopore R10.4.1 Flow Cells. Flow cell priming and sequencing reagents are also included as part of the sequencing kit and have been reformulated to be compatible with the Kit 14 chemistry upgrade and R10.4.1 nanopore. Other updates include higher capture rate of gDNA to enable lower flow cell loading amounts, and fuel fix technology, allowing users to run longer experiments without the need for fuel addition during the run.

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 24 V14 is supported by the EPI2ME wf-16S workflow, which can be used to analyse data from the 16S protocol. Deconvolution of barcoded sequencing data is supported by Dorado and EPI2ME which classify the barcode sequence and sort reads into corresponding folders.

The 16S Barcoding Kit allows multiplexing of ≤24 samples in a single run.