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 Rapid PCR Barcoding Kit offers the fastest and simplest method of preparation of barcoded libraries for low quantities of gDNA (1-5 ng), with only ~15 mins of hands-on preparation time.
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 12 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 features:
| Feature | Property |
|---|
| Preparation time | 15 minutes + PCR |
| Input requirement | 1-5 ng gDNA per sample |
| PCR Required | Yes |
| Fragmentation | Transposase-based |
| Read length | ~2 kb |
| Pack size | 6 reactions |
| Stability | Shipped 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. |
This kit is optimised to offer a simple workflow for those with a limited amount of input material. It has been observed that the fragment length produced after PCR is largely independent of the input fragment length, with the PCR products producing a peak at around 2 kb on an Agilent Bioanalyzer. This corresponds to a read-length of ~2 kb in nanopore sequencing. The length limit is due to the method of library preparation, rather than the processivity limit of the DNA polymerase.
Typical read-length histogram observed when preparing libraries with the Rapid PCR Barcoding Kit.
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 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), with only ~15 mins of hands-on preparation time.
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 12 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 contains 12 unique barcodes. There are sufficient barcodes and other reagents to generate six libraries with 12 barcoded samples in each one, which allows for processing a total of 72 samples.
| Name | Acronym | Cap colour | No. of vials | Fill volume per vial (µl) |
|---|
| Fragmentation Mix | FRM | Brown | 3 | 30 |
| Rapid Adapter | RAP | Green | 1 | 10 |
| Sequencing Buffer | SBQ | Red | 1 | 300 |
| Loading Beads | LB | Pink | 1 | 360 |
| Rapid Barcode Primer 1-12 | RLB 01-12 | Clear | 12 | 10 |
The Q-Line Flow Cell Priming Kit is also supplied.
| Name | Acronym | Cap colour | No. of vials | Fill volume per vial (µl_ |
|---|
| Flush Buffer | FB | Blue | 6 | 1,170 |
| Flush Tether | FLT | Purple | 1 | 200 |
Compatibility
The Q-Line Ligation Sequencing Kit can be used together with:
Kits
- Q-Line Flow Cell Wash Kit (Q-EXP-WSH004)
- Q-Line Flow Cell Priming Kit (Q-EXP-FLP002)
Flow cells
- Q-Line MinION GridION Flow Cell R9 version (Q-FLO-MIN106D)
Devices
- GridION Q Sequencing Device Mk1
