Description
Information
Rapid Barcoding Kit 96
This kit is recommended for users who:
- wish to multiplex samples to reduce price per sample
- need a PCR-free method of multiplexing to preserve additional information such as base modifications
- require a short preparation time
- have limited access to laboratory equipment.
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.
| Feature | Property |
| Preparation time | 60 minutes |
| Input requirement | 50 ng of high molecular weight gDNA (>30 kb) |
| PCR Required | No |
| Fragmentation | Transposase-based |
| Read length | Random distribution, dependent on input fragment length |
| Read type produced | 1D |
| Typical throughput* | ••• |
| Associated protocols | Rapid Barcoding Kit 96 (SQK-RBK110.96)
PCR tiling of SARS-CoV-2 virus with rapid barcoding (SQK-RBK110.96) |
| Multiplexing options | Rapid Barcoding Kit
Flow Cell Wash Kit |
| Pack size | 3 x 96 reactions with reagents to support 12 x 24 reactions |
| Stability | Shipped at 2–8° C
Long-term storage -20° C Oxford Nanopore Technologies deem the useful life of the product to be 3 months from receipt by the customer. |
The kit is optimised for simplicity and speed, rather than for obtaining maximum throughput. Due to the simple nature of the workflow and the fact that little sample manipulation is required, some very long reads can be achieved with this kit, despite the required transposase fragmentation. However, for long reads to be observed in sequencing, long fragments need to be present in the sample in the first place.
The workflow is PCR-free and therefore removes PCR bias and retains base modification information which can be analysed using tools developed in the Nanopore Community.
Deconvolution of barcoded sequencing data is supported by MinKNOW and Guppy software, which classify the barcode sequence and sort reads into corresponding folders.
Further considerations:
The Rapid Barcoding Kit recommends an input of 50 ng gDNA per sample, and is optimised for samples which contain long fragments (>30 kb). Addition of less than 50 ng, or shorter fragments could compromise sequencing throughput and read length. For samples where only lower inputs are possible, the Rapid PCR Barcoding Kit is available.
Shipping and logistics:
Flow cells and kits are shipped together at 2–8°C.
Upon receipt, please place the right product in the right long-term storage location.
Workflow
The Rapid Barcoding Kit 96 generates barcoded sequencing libraries from extracted gDNA in 60 minutes using a simple protocol. At the heart of the kit is a transposase which simultaneously cleaves template molecules and attaches barcoded tags to the cleaved ends: there are 12 unique barcoded tags in the kit. Barcoded samples are pooled, purified using SPRI beads, and Rapid Sequencing Adapters are then added to the tagged ends.
What’s in the box
Compatibility
The Rapid Barcoding Kit 96 can be used together with:
Kits
- Flow Cell Wash Kit (EXP-WSH004)
Flow cells
- FLO-MINSP6
- FLO-MIN106D
- FLO-MIN111
- FLO-PRO002
- FLO-FLG001
Devices
