Oxford Nanopore Technology (ONT) Ligation Sequencing library preparation with Native Barcode kit provides a median raw read accuracy of Q20+ (99%) with a protocol that is free from PCR, fragmentation, and size selection. There are numerous advantages to this approach, one of which is the elimination of PCR error and enzyme bias. Additionally, nanopore sequencing provides direct sequencing of native DNA molecules which can produce extremely long reads, as well as kinetic data for each molecule to infer base modifications of the nucleic acids.
Sequencing of native molecules does impose higher input requirements compared to PCR-based methods because the ligation-based preparation is less efficient. This effect is compounded further by the lack of amplification. Additionally, the nanopore platform is sensitive to certain compounds that can be carried over during extraction, such as polysaccharides (common for some fungi, plants, and insects) and excessive amounts of unprepared nucleic acids (e.g., excess RNA in a DNA library).
SeqCenter aims to provide nanopore sequencing in packages and prices that are applicable and affordable for projects of all sizes, including small genomes. By multiplexing all of our single sample packages, we are able to subdivide the output and cost of the nanopore flowcells for both our hybrid sequencing and ONT-only services.