Amplicon-seq of Puccinia graminis f. sp. tritici
Fungicide target genes (SdhA, SdhB, SdhC, SdhD, and Cyp51) of wheat pathogen Puccinia graminis f. sp. tritici was PCR amplified used primers described by Savva, Bryan et al. (Savva, L.; Bryan, A.; Vinopal, D.; Gonzalez-Navarro, O. E.; Kosgey, Z.; Ndungu, K. C.; Horo, J. T.; Danu, K. G.; Molla, M.; Alemayehu, Y.; et al. A portable, nanopore-based genotyping platform for near real-time detection of Puccinia graminis f. sp. tritici lineages and fungicide sensitivity). PCR products of these genes was pulled at equal concentrations, barcoded using  Rapid barcoding kit (Oxford Nanopore Technologies, Oxford, United Kingdom) and subjected to sequencing on the MiniON flow cell SQK-LSK114.