The Nanopore Learning course on “Human genome sequencing and analysis” has a video on Kit V14. Vania Costa, a Technical Services Specialist with Oxford Nanopore Technologies, described the accuracy of kit 14, which relies on R10.4.1 flow cells and pores. Costa explained that kit v14 has simplex Q20+ and duplex Q30+. Currently, there are ligation-based kits and rapid kits for kit 14. The ligation kit is optimized to deliver the highest output. The ligation kit blunts overheads and ligates sequencing adapters. The ligation sequencing kit can use amplicons and genomic DNA as templates. The rapid kits are transposase based that cleave and tag DNA. A sequencing adapter is added to the DNA ends. The ultra long sequencing kit requires a higher mass and can generate 4+ Mb reads! Costa noted that there are PCR and PCR-free kits. With PCR-free options, information about modifications can be obtained. PCR-based options are useful if starting material is limited. Multiplexing is an option, including the combination of PCR and native barcoding. End-to-end workflows include protocols and EPI2ME analyses. Costa explained that with PromethION flow cells, there are workflows for complete human DNA sequencing and analysis. Costa highlighted the ONT Protocol Libraries, Protocol Builder, Protocol Selector, and other resources available on the website. The Support page also has frequently asked questions (FAQs) that I have found useful when learning about new protocols and kits.
