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Rapid Barcoding Kit 24 V14
SQK-RBK114.24

Simple and rapid library preparation, with barcoding for up to 24 gDNA samples.

This is an Early Access product

Product lead time: 4 weeks

This uses our latest Kit 14 chemistry

$699.00
2Early Access
  • Access the latest products from Oxford Nanopore Technologies
  • Provide feedback into the innovation team as the product develops
  • Product Warranty may initially be limited to 1 month as it is so new
  • Product routinely iterates as new features and capabilities are added
  • Estimated lead time in store
  • Product subject to availability

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.

No barcodes6 barcodes12 barcodes24 barcodes
Flow cell price$500$500$500$500
Library price$99$99$99$99
Barcode price-$25$50$75
Price per sample$599$104$54$28

The Rapid Barcoding Kit 24 V14 features

FeatureProperty
Preparation time60 minutes
Input requirement•50 ng gDNA (>30 kb) per sample for >4 barcodes
•200 ng gDNA per sample for <4 barcodes
PCR RequiredNo
FragmentationTransposase-based
Read lengthRandom distribution, dependent on input fragment length
Kit chemistryKit 14 (V14)
Associated protocolsComing soon: Rapid Sequencing gDNA - barcoding Kit V14 (SQK-RBK114.24)
Pack size6 reactions
StabilityShipped 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 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 median 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.

Please 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

Coming soon: Rapid sequencing gDNA - barcoding kit V14 (SQK-RBK114.24)

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

What's in the box

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

RBK114.24 tubes

NameAcronymCap colourNo. of vialsFill volume per vial (µl)
Rapid AdapterRAGreen115
Adapter BufferADBClear1100
AMPure XP BeadsAXPAmber21200
Elution BufferEBBlack1500
Sequencing BufferSBRed1700
Library BeadsLIBPink1600
Library SolutionLISWhite cap, pink label1600
Flow Cell FlushFCFBlue61170
Flow Cell TetherFCTPurple1200
Rapid BarcodesRB01-24Clear2415

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 V14 (EXP-FLP004)
  • Control Expansion (EXP-CTL001)

Flow cells:

R10.4.1:

  • FLO-MIN114
  • FLO-PRO114M

Devices:

  • MinION Mk1B
  • MinION Mk1C
  • GridION
  • PromethION