Information
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
Please note that to use this kit, you will need to purchase additional 3rd party materials: see the "3rd party materials" tab for more detail.
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.
The Q-Line Rapid Barcoding Kit 96 V14 features:
| Feature | Property |
|---|
| Preparation time | <60 minutes |
| Input requirement | 50–200 ng gDNA (dependent on sample fragment length) |
| PCR required | No |
| Fragmentation | Transposase-based |
| Read length | Random distribution, dependent on sample fragment length |
| Kit chemistry | Kit 14 |
| Pack size | Up to 12 reactions
Options: 12 libraries containing 24 barcodes, 6 libraries containing 48 barcodes, 3 libraries containing 96 barcodes |
| Stability | Store at –30°C to –20°C until the labelled expiration date.
Q reagents are supplied with a minimum of 3 months shelf life remaining at the time of delivery.
Expiration dates are displayed in DD-MMM-YYYY format. |
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.
Deconvolution of barcoded sequencing data is supported by the sequencing software, which classify the barcode sequence and sort reads into corresponding folders.
Further considerations:
The Q-Line Rapid Barcoding Kit 96 V14 recommends an input of 50 ng high molecular weight gDNA (>30 kb) or 200 ng of shorter amplicon/plasmid DNA per sample.
Please note: <1 kb fragments could compromise sequencing throughput and read length.
Shipping and logistics:
Bulk order options are available. All orders can be split for delivery to best suit your needs.
Flow cells and kits are shipped together at 2°C to 8°C. The packaging is designed to protect the flow cells from freezing.
Workflow
The Q-Line Rapid Barcoding Kit 96 V14 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 96 unique barcoded tags in the kit. Barcoded samples are pooled, purified using AMPure XP beads, and Rapid Sequencing Adapters are then added to the tagged ends.
What's in the box
The Q-Line Rapid Barcoding Kit 96 V14 (Q-SQK-RBK114.96) contains the following reagents:
| Name | Abbreviation | Cap colour | Quantity | Minimum fill volume per container (µl) |
|---|
| Rapid Adapter | RA | Green | 2 | 9 |
| Adapter Buffer | ADB | Clear | 1 | 42 |
| AMPure XP Beads | AXP | Amber | 3 | 1,000 |
| Elution Buffer | EB | Black | 1 | 720 |
| Sequencing Buffer | SB | Red | 1 | 450 |
| Library Beads | LIB | Pink | 1 | 306 |
| Flow Cell Flush | FCF | N/A | 1 | 14,040 |
| Flow Cell Tether | FCT | Purple | 2 | 180 |
| Rapid Barcodes | RB01-96 | - | 3 plates | 1.5 µl per well |
Note: The AMPure XP reagent manufactured by Beckman Coulter Inc. can be stored at –30°C to –20°C with the kit, without detriment to reagent stability.
Compatibility
The Q-Line Rapid Barcoding Kit 96 V14 can be used together with:
Kits
- Q-Line Control Expansion Kit (Q-EXP-CTL001)
- Q-Line Flow Cell Wash Kit (Q-EXP-WSH004)
Flow cells
- Q-Line MinION Flow Cell R10 (Q-FLO-MIN114)
Devices
- GridION Q sequencing device (Q-GRD-MK1)
