In addition to simplex reads, V14 chemistry and R10.4.1 nanopore combination can also produce duplex reads to further increase accuracy. Duplex reads combine data from both strands into one basecall to generate single molecule accuracy to ~Q30 or higher. The duplex approach with V14 chemistry has already demonstrated perfect reads up to 72 kb in length and Q30 reads at 260 kb.

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Miten Jain's team at University of California, Santa Cruz, USA have been assessing recent improvements in read length, accuracy, and software for ultra-rapid, nanopore-based whole-genome sequencing analysis. Specifically focusing upon Kit 14 here, Miten presented data generated in the past few days prior to this talk, showing the generation of over 140 Gb of sequencing data in a single PromethION Flow Cell. This high output is achieved with lower DNA and library input requirements than for the previous chemistry. Also highlighted was a notable improvement in raw read accuracy, particularly evident in homopolymer regions where lengths of up to approximately 20 bases were accurately called.

The team is currently generating their first human genome assemblies using the latest nanopore chemistry and flow cells; however initial results using sheared genomic DNA, are reaching NG50 scores exceeding 20 Mb. They are also evaluating the use of duplex data and developing further variant calling and methylation analysis tools. With the developments presented here, Miten concluded that “phased, Q40+ accuracy de novo assemblies may be achievable”.