Ligation Sequencing Kits enable ligation-based, PCR-free nanopore sequencing of native DNA.
- Sequence native DNA — eliminating PCR bias and retaining 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). 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.
The Ligation Sequencing Kit 76487-140 is optimized to generate maximum throughput and read length. For highest data yields, we recommend starting with 50 to 100 fmol of pure input DNA. Starting with lower amounts of input material, or impure samples, can affect library preparation efficiency and can reduce sequencing throughput.
The Ligation Sequencing Kit 76487-138 is optimized to generate maximum throughput. For highest data yields, we recommend starting with 100 to 200 fmol of pure input DNA. Starting with lower amounts of input material, or impure samples, can affect library preparation efficiency and can reduce sequencing throughput.
Users can either start with 1 μg of gDNA, quantified using the Qubit fluorometer, or 50 fmol (76487-140)/100 to 200 fmol (76487-138) 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 neither advised nor required. Note: if long reads are required, then long fragments must be present in the starting material. 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 to 2.2.
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.
For sample multiplexing see expansion packs.
Informacje do zamówienia: 76487-138 [LSK109] will be discontinued three months after the introduction of the SQK-NBD110.24 (a specific Native Barcoding Expansion Pack for 76487-140 [LSK110]), which is anticipated in autumn 2021; however, use of 76487-138 [LSK109] for all COVID-related projects will be supported indefinitely.
Informacja o dostawie: Flow cells and kits are shipped together at 2 to 8 °C. 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.
Uwaga: Oxford Nanopore Technologies products are currently for research use only. For multiplexing samples using ligation-based sample preparation, use 76487-138 and relevant Native Barcoding Expansion Pack; for singleplex samples, use 76487-140.