Information
The Ligation Sequencing Kit V14 (SQK-LSK114) is highly recommended for users who:
- Want to achieve modal raw read accuracy of Q20+ (99%) and above
- Want to obtain duplex data
- Want to optimise their sequencing experiment for accuracy and output
- Require control over read length
- Would like to utilise upstream processes such as size selection or whole genome amplification
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.
Also included are the MinION Flow Cells (R10.4.1) The R10 series of nanopores contains a double reader-head, and is suitable for experiments where high consensus accuracy is required. Paired with the Kit 14 chemistry, R10.4.1 generates data at a modal accuracy above 99%.
The Ligation Sequencing Kit V14 features:
The Ligation Sequencing Kit V14 offers a flexible method of preparing sequencing libraries from dsDNA (e.g. gDNA, cDNA or amplicons). The library preparation method is straightforward: DNA ends are repaired and dA-tailed using the NEBNext End Repair/dA-tailing module before the sequencing adapters, supplied in the kit, are ligated onto the prepared ends.
The kit is optimised to achieve sequencing accuracies of over 99% (Q20+) with high output on our latest nanopore: R10.4.1. Our flow cell priming and sequencing reagents have been reformulated to be compatible with this improved Kit 14 adapter and R10.4.1 nanopore.
Kit 14 also includes previous updates such as the 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.
Users can either start with 1 μg of gDNA or 100-200 fmol of shorter-fragment input such as amplicons or cDNA. If your experiment requires long reads, it is recommended to start with full-length gDNA, and fragmentation/shearing is not advised. Please note: if long reads are required, high molecular weight DNA should be used as input. To determine purity, we suggest using the Nanodrop to measure the A260/280 and A260/230 ratios and we recommend that the sample should meet the following criteria:
- A260/280 = 1.8
- A260/230 = 2.0-2.2
The Ligation Sequencing Kit V14 is compatible with upstream processes such as target enrichment by sequence capture, whole genome amplification (for applications where under 1 ng of sample is available) and size selection (for enrichment of specified fragment lengths, using our Short Fragment Eliminator kit (EXP-SFE001)).
A) Read lengths observed when HMW gDNA was prepared using the Ligation Sequencing Kit with and without size selection (EXP-SFE001 treatment). B) Read N50 observed under the same sequencing conditions, demonstrating significant increase when size selected using EXP-SFE001.
PCR- and WGA-free workflows remove amplification bias and retain base modification information, which can be analysed using tools supported by Oxford Nanopore.
Further considerations:
Starting with lower amounts of input material, or impure samples, can affect library preparation efficiency and can reduce sequencing output. PCR can be used to generate more input material in cases where sample amount is limiting.
Shipping and logistics:
Flow cells and kits are shipped together at 2–8°C. The packaging is designed to protect the flow cells from freezing. Flow cells are stable at ambient temperatures and stored refrigerated for longer periods. Sequencing kits are stable at room temperature for up to seven days, so users should not be concerned about their products should they arrive at ambient temperature.
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.
Storage and stability:
Please store your flow cells as follows:
- Room temperature: flow cells can be stored, unopened, at room temperature for one month.
- 2-8°C: flow cells can be stored, unopened, at 2-8°C for 12 weeks.
We recommend you check your flow cell before preparing your DNA/RNA library as part of the warranty process.
Flow cell returns:
The flow cell pouch provided serves as the flow cells returns packaging. Flow cells contain finite materials which, when returned to Oxford Nanopore, are responsibly recycled.
More information on how to carry out a return can be found on the Nanopore Community.
Workflow
The Ligation Sequencing Kit offers a flexible method of preparing sequencing libraries from dsDNA (e.g. gDNA, cDNA or amplicons). The library preparation method is straightforward: DNA ends are FFPE repaired and end-prepped/dA-tailed using the NEBNext End Repair/dA-tailing module, and then sequencing adapters, supplied in the kit, are ligated onto the prepared ends.
Safety and legal
English Safety Data Sheet for Ligation Sequencing Kit V14 (SQK-LSK114)
Spanish Safety Data Sheet for Ligation Sequencing Kit V14 (SQK-LSK114)
English Safety Data Sheet for MinION Flow Cell (FLO-MIN114)
Spanish Safety Data Sheet for MinION Flow Cell (FLO-MIN114)
Customers are required to perform a flow cell check within 12 weeks of purchase. Oxford Nanopore Technologies will replace any flow cell with fewer than 800 nanopores, when the result is reported within two days of performing the flow cell check, and when the storage recommendations have been followed. The warranty on this product is 12 weeks from receipt by the customer.
There is a 9-month contract length available for all consumable purchases. For more information, please see the Nanopore Product Terms and Conditions.
What's in the box
This promo pack contains:
- 10 x MinION/GridION Flow Cells (FLO-MIN114)
- 2 x Ligation Sequencing Kit V14 (SQK-LSK114)