Ligation Sequencing Kits enable ligation-based, PCR-free nanopore sequencing of native DNA.
- Sequence native DNA — eliminate PCR bias and retain base modifications
- High sequencing yields
- Control over read length
- Suitable for upstream processes such as size selection or whole-genome amplification
Ligation Sequencing Kits offer a flexible method of preparing sequencing libraries from dsDNA (e.g. gDNA, cDNA, or amplicons) for nanopore sequencing. A range of Ligation Sequencing Kits are available to suit your experimental requirements:
Ligation Sequencing Kit (SQK-LSK112) – the latest nanopore sequencing chemistry, optimised to achieve sequencing accuracies of over 99% (Q20+) when combined with R10.4 flow cells (FLO-MIN112). This kit is available in three formats: Ligation Sequencing Kit (SQK-LSK112) – for singleplex samples, Native Barcoding Kit 24 (SQK-NBD112.24) – standalone kit enabling PCR-free multiplexing of up to 24 samples per run and Native Barcoding Kit 96 (SQK-NBD112.96) – standalone kit enabling PCR-free multiplexing of up to 96 samples per run.
Ligation Sequencing Kit (SQK-LSK110) – optimised for maximum data yield from singleplex samples. For PCR-free sample multiplexing, see Native Barcoding Kits SQK-NBD112.24 or SQK-NBD112.96 on this page. For PCR-based sample multiplexing, see PCR Barcoding Expansion 1-12 and 1-96.
Ligation Sequencing Kit (SQK-LSK109) – this product is anticipated to be discontinued in early 2022 except where used in COVID-related projects, which will be supported indefinitely.
The library preparation method is straightforward: DNA ends are repaired and dA-tailed using the NEBNext End Repair/dA-tailing module, and then sequencing adapters, supplied in the kit, are ligated onto the prepared ends. With no requirement for amplification, these kits eliminate potential PCR bias and preserve base modifications, which can be detected alongside the nucleotide sequence.
For highest data yields, we recommend starting with 50 to 100 fmol* of pure input DNA for singleplex samples. Starting with lower amounts of input material, or impure samples, can affect library preparation efficiency and can reduce sequencing throughput.
*100 to 200 fmol for Ligation Sequencing Kit (SQK-LSK109).
Ligation Sequencing Kits are 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 the BluePippin, for example). When size selecting, we recommend increasing the amount of input used, as size selection can be a wasteful process.
The Ligation Sequencing Kit (SQK-LSK112) and the Native Barcoding Kits (SQK-NBD112.24 / SQK-NBD112.96) are compatible with both R10.4 and R9.4.1 Flow Cells. Ligation Sequencing Kits (SQK-LSK110 and SQK-LSK109) are compatible with R9.4.1 Flow Cells.
Ordering information: SQK-LSK109 is anticipated to be discontinued in early 2022 except where used in COVID-related projects, which will be supported indefinitely.
Delivery information: Flow cells and kits are shipped together 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.