The Nanopore Learning course on Human genome sequencing and analysis has a video on the Kit V14 performance. Vania Costa, a member of Oxford Nanopore Technologies Technical Services Team, explained how kit 14 uses R10.4.1 flow cells for increased simplex Q20+ accuracy and duplex Q30+ accuracy. The R9 kit will be discontinued at the end of 2023. Kit 14 duplex is described as “reading both strands independent of read length.” Duplex reads can be kilobases long. Washing a flow cell can maximize yield. Costa shared data with kit washing at 14 and 40 hours. ONT recommends preparing a new library for loading after washing, though recovering a library is possible. A library can be recovered and loaded on up to three flow cells sequentially. Kit 14 offers highly accurate genetic and epigenetic information with the super accuracy basecalling model (SUP). With long and ultra-long reads, long-range phasing can be achieved. Costa suggested using Flye for assembly and one round of polishing with Medaka. Today, we are using the kit 14 rapid barcoding kit and a R10.4.1 flow cell from December. I hope that the yield for whole genome sequencing of microbiomes is enough for class analyses.
