Description
Information
Ultra-Long DNA Sequencing Kit
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
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 using a Circulomics 5 mm Nanobind disk to remove free adapter and short DNA fragments.
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.
Users have achieved the following from an ultra-long preparation for both MinION and PromethION R9.4.1 flow cells is as follows:
| Feature | Property | ||
| Preparation time | ~610 minutes | ||
| Input requirement |
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| PCR Required | No | ||
| Fragmentation | Transposase-based | ||
| Read length | R9.4.1 flow cells: 50–100+ kb N50, reads up to 4+ Mb |
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| Read type produced | 1D | ||
| Typical throughput* |
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| Associated protocols | Ultra-Long DNA 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 at 2–8°C.
Upon receipt, please place the right product in the right long-term storage location.
Workflow
The Ultra-Long DNA Sequencing Kit generates sequencing libraries from extracted ultra-high molecular weight gDNA in ~610 minutes. At the heart of the 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, for example using a Circulomics 5 mm Nanobind disk, to remove free adapter and short DNA fragments.
What’s in the box
The Ultra-Long DNA Sequencing Kit contains sufficient reagents to generate sequencing libraries for six reactions, where each reaction supports three separate loads of PromethION Flow Cells and six MinION Flow Cells.
| Name | Acronym | Cap colour | No. of tubes | Fill volume (μl) |
| Rapid Adapter | RAP F | Green | 1 | 30 |
| Fragmentation Mix | FRA | Brown | 1 | 36 |
| FRA Dilution Buffer | FDB | Clear | 1 | 1464 |
| Elution Buffer | EB | Black | 1 | 1350 |
| Sequencing Buffer | SQB | Red | 1 | 1350 |
| Priming Tether | PT | Purple | 1 | 540 |
| Flush Buffer | FB | White cap, light blue stripe on label | 3 | 7020 |
| L Fragment Buffer | LFB | White cap, orange stripe on label | 2 | 6000 |
Compatibility
The Ultra-Long DNA Sequencing Kit can be used together with:
Kits
- Flow Cell Wash Kit (EXP-WSH004)
Flow cells
- FLO-MINSP6
- FLO-MIN106D
- FLO-MIN111
- FLO-PRO002
Devices
