Microbial Amplicon Barcoding Kit 24 V14

Amplification and full-length sequencing of 16S rRNA for bacterial profiling and ITS for fungal profiling, with barcoding for up to 24 samples.

• Enables full-length sequencing of 16S rRNA (bacterial profiling) and ITS (fungal profiling)
• Provides genus-level bacterial identification
• Delivers a rapid, cost-effective workflow - faster than ligation-based approaches
• Utilises a fragmentation-free process that preserves full-length amplicons

The microbial amplicon barcoding sequencing kit 24 V14 offers a method of amplifying the entire 16S rRNA or ITS gene from extracted gDNA, followed by library preparation. This kit contains updated primers that enable full-length sequencing of 16S rRNA for bacterial profiling and ITS for fungal profiling from diverse sample types.

Please note: These primer schemes are aligned with industry standard and provide coverage for the entire 16S rRNA and ITS rRNA gene respectively. However, the primers used bind within regions that are not 100% conserved across species and therefore it is unable to guarantee amplification for all species. Please ensure to run a control experiment to determine if targeted species will be amplified using the primers provided in this kit.

The DNA is amplified by PCR using specific 16S or ITS primers. Up to 24 amplicon samples are then barcoded. These barcodes contain 5’ tags which facilitate the ligase-free attachment of rapid sequencing adapters. 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.

This kit uses Kit 14 chemistry, which includes improved modal raw read sequencing accuracies with high output on the latest nanopore: R10.4.1. The flow cell priming and sequencing reagents are also included as part of the sequencing kit and are compatible with the R10.4.1 nanopore. This chemistry presents a 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.

The kit is supported by the EPI2ME wf-16S workflow, which can be used to analyse data from the microbial amplicon barcoding sequencing kit 24 V14 (SQK-MAB114.24) protocol. Deconvolution of barcoded sequencing data is supported by Dorado and EPI2ME which classify the barcode sequence and sort reads into corresponding folders.

Sample multiplexing is ≤24.

 : Kits are shipped at 2 to 8 °C in an environmentally friendly temperature-controlled shipping box. Upon receipt, please place the product in a long-term storage location at ‒20 °C.

 : Oxford Nanopore Technologies products are not intended for use for health assessment or to diagnose, treat, mitigate, cure, or prevent any disease or condition.