Information
This kit is recommended for users who:
- Are working with ultra high molecular weight DNA and would like read lengths with an N50 >50 kb
- Require an end-to-end workflow to sequence ultra high molecular weight DNA
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.
This is a Legacy product
This kit is soon to be discontinued and we recommend all customers to upgrade to the latest chemistry for their relevant kit which is available on the Store. If customers require further support for any ongoing critical experiments using a Legacy product, please contact Customer Support via email: support@nanoporetech.com.
The Ultra-Long DNA Sequencing Kit offers a means of preparing ultra high molecular weight (uHMW) DNA for sequencing, which has shown to give N50s >50 kb and reads up to 4+ Mb. The kit is based on transposase chemistry: the transposase simultaneously cleaves template molecules and attaches tags to the cleaved ends. Rapid Sequencing Adapters are then added to the tagged ends. The last step is an overnight elution of the DNA library.
The kit includes a new type of Rapid Adapter (RAP F), which is designed such that its consumption of fuel during a sequencing run is reduced significantly (over RAP), resulting in much better maintenance of enzyme speed. As such, the new adapter results in higher sequencing output.
For an end-to-end workflow, we have used the NEB Monarch® HMW DNA Extraction Kit for Tissue to extract uHMW gDNA, enabling users to extract and sequence their sample in just one day.
Validated inputs for the end-to-end protocol are:
- 6 million human cells isolated from culture (e.g. GM24385)
- 6 million peripheral blood mononuclear cells (PBMCs) from blood
These inputs produce high read N50s and, when paired with the Flow Cell Wash Kit (EXP-WSH004), deliver high outputs on both MinION and PromethION.
The Ultra-Long DNA Sequencing Kit features:
| Feature | Property |
|---|
| Preparation time | ~200 minutes + overnight elution (≥ 12 hours) |
| Input requirement | • Tissue culture cells (e.g. GM24385): 6 million
• Peripheral blood mononuclear cells: 6 million
|
| PCR Required | No |
| Fragmentation | Transposase-based |
| Kit chemistry | Kit 10 |
| Read length | R9.4.1 flow cells: 50–100+ kb N50, reads up to 4+ Mb |
| Associated protocols | Ultra-Long Sequencing Kit (SQK-ULK001) |
| Multiplexing options | Flow Cell Wash Kit |
| Pack size | Six reactions where each reaction supports three separate loads of either PromethION Flow Cells or MinION Flow Cells |
| 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. |
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 protocol outlines a complete workflow from extracting gDNA from cell culture or purified cells from whole blood using the NEB Monarch® HMW DNA Extraction Kit for Tissue to the sequencing of uHMW gDNA using the Ultra-Long DNA Sequencing Kit in a day.
At the heart of the sequencing kit is a transposase which simultaneously cleaves template molecules and attaches tags to the cleaved ends. Rapid Sequencing Adapters are then added to the tagged ends, followed by an overnight elution before loading the DNA library onto a flow cell and sequencing.
O/N = Overnight elution ≥ 12 hours
Protocols
Rapid sequencing gDNA - Ultra-Long Sequencing Kit (SQK-ULK001)
To access the protocol, you will need to register for a Nanopore Community account.
What's in the box
The Ultra-Long DNA Sequencing Kit contains sufficient reagents to generate six reactions, where each reaction supports three separate loads of either PromethION Flow Cells or MinION Flow Cells.
| Name | Acronym | Cap colour | No. of tubes | Fill volume per vial (µl) |
|---|
| Rapid Adapter | RAP F | Green | 1 | 40 |
| Fragmentation Mix | FRA | Brown | 1 | 50 |
| FRA Dilution Buffer | FDB | Clear | 1 | 1,600 |
| Elution Buffer | EB | Black | 1 | 1,500 |
| Sequencing Buffer | SQB | Red | 1 | 1,500 |
| Priming Tether | PT | Purple | 1 | 600 |
| Flush Buffer | FB | White cap, light blue stripe on label | 3 | 7,200 |
| L Fragment Buffer | LFB | White cap, orange stripe on label | 2 | 6,500 |
Compatibility
The Ultra-Long DNA Sequencing Kit can be used together with:
Kits
- Flow Cell Wash Kit (EXP-WSH004)
Flow cells
Devices
- MinION Mk1B
- MinION Mk1C
- GridION
- PromethION