期刊
VIRUSES-BASEL
卷 13, 期 8, 页码 -出版社
MDPI
DOI: 10.3390/v13081424
关键词
biosecurity; ONT; MinION; diagnostics; plant viruses; high-throughput sequencing; rRNA depletion
类别
资金
- MPI
The adoption of Oxford Nanopore Technologies (ONT) sequencing in plant virology has been slow, but its promise of real-time generation of long nucleotide sequences makes it an attractive method for both low- and high-scale virus diagnostics. The portability of the MinION and Flongle platforms, combined with lower costs and improved read accuracy, highlight ONT's feasibility as a valuable component to diagnostician's toolkit.
The adoption of Oxford Nanopore Technologies (ONT) sequencing as a tool in plant virology has been relatively slow despite its promise in more recent years to yield large quantities of long nucleotide sequences in real time without the need for prior amplification. The portability of the MinION and Flongle platforms combined with lowering costs and continued improvements in read accuracy make ONT an attractive method for both low- and high-scale virus diagnostics. Here, we provide a detailed step-by-step protocol using the ONT Flongle platform that we have developed for the routine application on a range of symptomatic post-entry quarantine and domestic surveillance plant samples. The aim of this methods paper is to highlight ONT's feasibility as a valuable component to the diagnostician's toolkit and to hopefully stimulate other laboratories towards the eventual goal of integrating high-throughput sequencing technologies as validated plant virus diagnostic methods in their own right.
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