S. aureus MW2 WT ONT Direct RNA seq
Ribosomal RNAs were removed from 10g of total RNA with the RiboZero Bacteria kit (Illumina, CA). Enriched mRNAs were treated with E. coli Poly(A) Polymerase (NEB) in the presence of 10mM ATP to add a polyA tail. PolyAdenylated mRNAs were converted into a nanopore-compatible RNA library with the Direct RNA Sequencing kit from Oxford Nanopore (SQC-RNA002). The library was sequenced on an R9.4 flowcell in a GridION instrument (Oxford Nanopore) for 24hs and basecalled in real-time in the instrument with the software Guppy.