Information

This kit is recommended for users who:

• Wish to multiplex up to 24 samples to reduce price per sample
• Require a short preparation time
• Need a PCR-free method of multiplexing to preserve additional information such as base modifications
• Have limited access to laboratory equipment

This is an Early Access product.

For more information about our Early Access programmes, please see this article on product release phases.

Please note that to use this kit, you will need to purchase additional 3rd party reagents: see the «3rd Party Materials» tab for more detail.

Barcoding or multiplexing is useful when the amount of data required per sample is less that 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.

Multiplexing samples onto one flow cell can reduce the cost per sample for a user. Below are some worked examples. Please note, this is an example and pricing depends on the kit purchased.

The Rapid Barcoding Kit 24 V14 features

The Rapid Barcoding Kit 24 V14 (SQK-RBK114.24) is a standalone kit optimised for simplicity and speed to sequence up to 24 libraries using a PCR-free method. The kit uses a transposase to simultaneously cleave template molecules and attach barcoded tags to the cleaved ends. There are 24 unique barcodes provided in this kit.

This kit has been updated to use our newest Kit 14 chemistry which includes improved modal raw read sequencing accuracies with higher output on our latest nanopore: R10.4.1. The 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.

Kit 14 also includes previous updates such as the higher capture rate of DNA 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.

Note: This kit is able to generate high sequencing accuracies of over 99% (Q20+) and duplex data but has been optimised for speed and simplicity. If users require high accuracy, we recommend using our Native Barcoding Kit 24 V14 kit (SQK-NBD114.24).

Due to the simple nature of the workflow and the fact that little sample manipulation is required (e.g. minimal pipetting steps and no clean-up steps) some very long reads can be achieved with this kit, despite the required transposase fragmentation. However, in order 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, removing the PCR bias and retains information about base modifications, which can be analysed using tools developed in the Nanopore Community.

Further considerations:

The Rapid Sequencing Kit recommends an input of 50 ng gDNA, and is optimised for samples which contain long fragments (>30 kb). Addition of less than 50 ng, or the presence of shorter fragments could compromise sequencing output and read length.

Shipping and logistics

Flow cells and kits are shipped together in an environmentally friendly temperature-controlled shipping box.

Products are shipped to customers within the USA and EU Monday to Thursday. Shipments to Canada, Norway, Korea and Japan are expedited Monday to Wednesday; with Australia and New Zealand leaving our warehouses on a Friday. Shipments to the rest of the world are made on Mondays to allow the full working week for packages to arrive.

The delivery charges are calculated when a quote is raised or during checkout. Once an order is made, the delivery ID and delivery information can be tracked in the Store.

Workflow

The Rapid Barcoding Kit 24 V14 generates sequencing libraries from extracted gDNA using a transposase which simultaneously cleaves template molecules and attaches barcoded tags to the cleaved ends. The barcoded samples are then SPRI cleaned and pooled before adding the Rapid Sequencing Adapters to the tagged ends.

Protocols

• Rapid sequencing DNA V14 – barcoding (SQK-RBK114.24 or SQK-RBK114.96)

• Rapid Sequencing V14 – Plasmid sequencing (SQK-RBK114.24 or SQK-RBK114.96)

To access the protocols, you will need to register for a Nanopore Community account.

Safety and legal

English Safety Data Sheet for Rapid Barcoding Kit 24 V14 (SQK-RBK114.24)

Spanish Safety Data Sheet for Rapid Sequencing Kit 24 V14 (SQK-RBK114.24)

What’s in the box

The Rapid Barcoding Kit 24 V14 (SQK-RBK114.24) contains enough reagents for six reactions.

This Product Contains AMPure XP Reagent Manufactured by Beckman Coulter, Inc. and can be stored at -20°C with the kit without detriment to reagent stability.

3rd party materials

Consumables

• 1.5 ml Eppendorf DNA LoBind tubes
• 2 ml Eppendorf DNA LoBind tubes
• 0.2 ml thin-walled PCR tubes
• Eppendorf twin.tec® PCR plate 96 LoBind, semi-skirted (Cat # 0030129504) with heat seals
• Nuclease-free water (e.g. ThermoFisher, cat # AM9937)
• Freshly-prepared 80% ethanol in nuclease-free water
• (Optional) Bovine Serum Albumin (BSA) (50 mg/ml) (e.g Invitrogen™ UltraPure™ BSA 50 mg/ml, cat# AM2616)

Equipment

• Ice bucket with ice
• Microplate centrifuge, e.g. Fisherbrand™ Mini Plate Spinner Centrifuge (Fisher Scientific, #11766427)
• Timer
• Thermal cycler or heat block at 30°C and 80°C
• Magnetic rack
• Hula mixer (gentle rotator mixer)
• P1000 pipette and tips
• P200 pipette and tips
• P100 pipette and tips
• P20 pipette and tips
• P2 pipette and tips
• Multichannel pipette

Optional equipment

• Qubit fluorometer (or equivalent for QC check)

Compatibility

The Rapid Barcoding Kit 24 V14 (SQK-RBK114.24) can be used together with:

Kits:

• Sequencing Auxiliary Kit V14 (EXP-AUX003)
• Flow Cell Wash Kit (EXP-WSH004)
• Flow Cell Priming Kit (EXP-FLP004)
• Flow Cell Priming Kit XL (EXP-FLP004-XL)
• Control Expansion (EXP-CTL001)
• Rapid Adapter Auxiliary V14 (EXP-RAA114)

Flow cells:

R10.4.1:

• FLO-MIN114
• FLO-PRO114M
• FLO-FLG114

Devices:

• MinION Mk1B
• MinION Mk1C
• GridION
• PromethION