An overnight method to prepare ultra-high molecular weight DNA for sequencing ultra-long reads (N50 >50 kb).
- End-to-end workflow to sequence ultra-high molecular weight DNA
- Read lengths with an N50 >50 kb
The Ultra-Long DNA Sequencing Kit V14 offers a means of preparing ultra-high molecular weight (uHMW) DNA for sequencing, which has shown to give N50s >50 kb and read lengths 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. This kit utilises our latest Kit 14 chemistry, which includes improved median raw read sequencing accuracies with higher output on our latest R10.4.1 Flow Cells.
The Kit 14 chemistry also includes previous chemistry updates such as higher capture rate of DNA to enable lower flow cell loading amounts, and fuel fix technology, allowing users to run longer experiments without the need for fuel addition during the run.
Specific to the Ultra-Long DNA Sequencing Kit V14, we now supply the Extraction EB (EEB) and Precipitation Buffer (PTB) prepared for customer use and an updated Flush Tether UL (FTU) for Kit 14 compatibility. A Precipitation Star (PS) is also provided in this kit to aid DNA precipitation and recovery.
The end-to-end kit protocol, which has been developed using the NEB Monarch® HMW DNA Extraction Kit for Tissue, allows users to extract and sequence their sample in just one day. Alternative kits for generating uHMW are available but have not been validated by Oxford Nanopore Technologies.
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.
Delivery information: Kits are shipped at 2 to 8 °C in an environmentally-friendly temperature-controlled shipping box. Upon receipt, please place the product in a long-term storage location at -20 °C. Oxford Nanopore Technologies deem the useful life of the product to be 3 months from receipt by the customer.
Caution: Oxford Nanopore Technologies products are not intended for use for health assessment or to diagnose, treat, mitigate, cure, or prevent any disease or condition.
