Information
This kit is recommended for users who:
- Wish to multiplex up to 96 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
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 barcodes | 24 barcodes | 48 barcodes | 96 barcodes |
|---|
| Flow cell price | $500 | $500 | $500 | $500 |
| Library price | $99 | $99 | $99 | $99 |
| Barcode price | - | $75 | $150 | $300 |
| Price per sample | $599 | $28 | $15 | $9.36 |
The Rapid Barcoding Kit 96 V14 features
The Rapid Barcoding Kit 96 V14 (SQK-RBK114.96) is a standalone kit optimised for simplicity and speed to sequence up to 96 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 96 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 96 V14 kit (SQK-NBD114.96).
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 200 ng gDNA. Addition of less than 200 ng 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 96 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.
What's in the box
The Rapid Barcoding Kit 96 V14 (SQK-RBK114.96) contains enough reagents for up to 12 reactions.
| Name | Acronym | Cap colour | No. of vials | Fill volume per vial (µl) |
|---|
| Rapid Adapter | RA | Green | 2 | 15 |
| Adapter Buffer | ADB | Clear | 1 | 100 |
| AMPure XP Beads | AXP | Amber | 3 | 1,200 |
| Elution Buffer | EB | Black | 1 | 1,500 |
| Sequencing Buffer | SB | Red | 1 | 1,700 |
| Library Beads | LIB | Pink | 1 | 1,800 |
| Library Solution | LIS | White cap, pink label | 1 | 1,800 |
| Flow Cell Flush | FCF | Clear cap, light blue label | 1 | 15,500 |
| Flow Cell Tether | FCT | Purple | 2 | 200 |
| Rapid Barcodes | RB01-96 | - | 3 plates | 8 µl per well |
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.
Compatibility
The Rapid Barcoding Kit 96 V14 (SQK-RBK114.96) can be used together with:
Kits:
- Sequencing Auxiliary Kit V14 (EXP-AUX003)
- Flow Cell Wash Kit (EXP-WSH004)
- Control Expansion (EXP-CTL001)
- Flow Cell Priming Kit (EXP-FLP004)
- Flow Cell Priming Kit XL (EXP-FLP004-XL)
- Rapid Adapter Auxiliary V14 (EXP-RAA114)
- Midnight RT PCR Expansion (EXP-MRT001)
Flow cells:
- FLO-MIN114
- FLO-PRO114M
- FLO-FLG114
Devices:
- MinION Mk1B
- MinION Mk1C
- GridION
- PromethION