Tonight I started the Human Genome Sequencing and Analysis Nanopore Learning course. Bala Periaswamy from ONT spoke about using Nanopore sequencing for human genomics. They noted that the long reads can be used to sequence larger portions to analyze human genome variations and epigenomics. They noted that kit 14 along with flow cells and motor proteins help improve quality of sequencing. A single PromethION flow cell can produce in excess of 100 Gbases. Data include methylation and typically allow for 30X coverage of a human genome. Periaswamy explained that Nanopore provides an “End-to-End protocol for Sequencing a Human Sample.” There are two protocols on the Nanopore website. The Ligation Sequencing gDNA v14 on PromethION flow cells can be used for 10 kb or 30 kb protocols. The protocols take ~220 and 290 mins, respectively. The N50 of 30 kb protocol does not have a fragmentation step. The Ultra-Long DNA Sequencing Kit v14 can produce and N50 and is compatible with R10.4.1 flow cells. The extraction recommends using the NEB Monarch HMW kit. The library preparation uses the rapid transposome complex. Nanopore recommends washing and reloading library to reduce pore blocking. The T2T Consortium used this protocol and has documentation on their website. Finally, Periaswamy recommended using EPI2ME and EPI2ME Labs and the human genome variation workflows. I think that learning about human DNA sequencing with Nanopore will help us improve our sequencing runs for non-bacterial DNA samples.
