Information
This kit is recommended for users who:
- Wish to multiplex samples to reduce price per sample
- Have a low starting amount of DNA
- Require a simple library preparation procedure
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 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 PCR Barcoding Kit offers the fastest and simplest method of preparation of barcoded libraries for low quantities of gDNA (1–5 ng).
At the heart of the kit is a transposase which simultaneously cleaves template molecules in each sample and attaches tags, which contain primer binding sites, to the cleaved ends. The kit contains 24 primers which are then used to amplify each sample. Each primer contains a barcode and 5’ tag which facilitates the ligase-free attachment of Rapid Sequencing Adapters. Amplified and barcoded samples are then pooled together and Rapid Sequencing Adapters are added to the pooled mix.
The Q-Line Rapid PCR Barcoding Kit 24 V14 features:
| Feature | Property |
|---|
| Preparation time | 60 minutes + PCR |
| Input requirement | 1–5 ng gDNA per sample |
| PCR required | Yes |
| Fragmentation | Transposase-based |
| Read length | Random distribution dependent on input fragment length |
| Kit chemistry | Kit 14 |
| Pack size | 6 reactions |
| Multiplexing options | Up to 24 samples per run |
| 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. |
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 PCR Barcoding Kit 24 V14 offers the fastest and simplest method of preparation of barcoded libraries for low quantities of gDNA (1–5 ng).
At the heart of the kit is a transposase which simultaneously cleaves template molecules in each sample and attaches tags, which contain primer binding sites, to the cleaved ends. The kit contains 24 primers which are then used to amplify each sample. Each primer contains a barcode and 5’ tag which facilitates the ligase-free attachment of Rapid Sequencing Adapters. Amplified and barcoded samples are then pooled together and Rapid Sequencing Adapters are added to the pooled mix.
What's in the box
The Q-Line Rapid PCR Barcoding Kit 24 V14 contains 24 unique barcodes and the following reagents:
| Name | Abbreviation | Cap colour | Quantity | Minimum fill volume per container (µl) |
|---|
| Fragmentation Mix | FRM | Amber | 1 | 144 |
| Rapid Adapter | RA | Green | 1 | 9 |
| Adapter Buffer | ADB | Clear | 1 | 21 |
| AMPure XP Beads | AXP | N/A | 1 | 4,580 |
| Elution Buffer | EB | Black | 1 | 90 |
| EDTA | EDTA | Blue | 1 | 576 |
| Sequencing Buffer | SB | Red | 1 | 225 |
| Library Beads | LIB | Pink | 1 | 153 |
| Flow Cell Flush | FCF | N/A | 1 | 7,020 |
| Flow Cell Tether | FCT | Purple | 1 | 180 |
| Rapid Barcode Primers (01-24) | RLB01-24 | - | 2 plates, 3 sets of primer barcodes per plate | 10 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 PCR Barcoding Kit 24 V14 can be used together with:
Kits
- 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)
