Nanopore-Seq of Lycaeides melissa: adult female thorax
we first repaired DNA molecules with the NEBNext FFPE DNA Repair Mix (NEB M6630) and NEBNext Ultra II end repair/dA-tailing Module (NEB E7546). This was done using 0.5-1.0 g genomic DNA per sample and in accordance with the manufacturers protocol. Individuals with similar DNA concentrations were barcoded using the Native Barcoding Expansion Pack 1-12 (EXP-NBD 104) and subsequently pooled to reach a total amount of 1 g of genomic DNA or a total number of 12 individuals (whichever came first); the number of individuals per pooled library was between 1 and 12 (Table S2). We then ligated adaptor oligos for sequencing onto the barcoded fragments in each pooled library using the Ligation Sequencing kit (SQK-LSK109). The ligation product from each library was then loaded onto R9.4 flow cell (FLO-Min106, ONT) for a sequencing run of a full 72 hours on a MinION.