title: multiplex PCR amplicon sequencing of TiLV_B1-2
design: Two sets of multiplex RT-PCR products were combined as a template for library preparation using the ligation sequencing kit (SQK-LSK109) and the native barcoding expansion 1-12 kit (EXP-NBD104) were used according to the Oxford Nanopore Technologies (ONT) standard protocols adapted for the Flongle flow cell. We used 250 ng PCR products for each sample (BCxx-BCxx), one unique native barcode (BC) per sample and washed the library of pooled barcoded samples with the Short Fragment Buffer (SFB) just before the elution step at the end of the protocol. DNA concentrations were determined between each step using the Qubit assay. The prepared TiLV library was loaded as per the standard protocol onto a Flongle flow cell (FLO-FLG106) for multiplex RT-PCR. Flongle flow cell was fitted to a Flongle adapter (FLGIntSP) for MinION.