Bulk RNA-seq of rattus norvegicus: sinonasal mucosa
long-read transcriptomics was performed using the direct cDNA sequencing kit (Oxford Nanopore Technologies, SQK-DCS109) according to the manufacturers recommendations with the following minor modification: the protocol was started with 200 ng poly(A) enriched RNA. In short, after first-strand cDNAs were synthesised using the Maxima H Minus Reverse Transcriptase (Thermo Fisher Scientific, EP0752), the RNA was degraded with RNase Cocktail Enzyme Mix (Thermo Fisher Scientific, AM2286) and second-strand cDNAs were synthesised using the LongAmp Taq 2X Master Mix (New England Biolabs, M0287L). The double-stranded cDNAs were end-repaired using the NEBNext Ultra II End Repair/dA-Tailing Module (New England Biolabs, E7546S). Sequencing adapters were ligated to the end-repaired strands using Blunt/TA Ligase Master Mix (New England Biolabs, M0367L). The cDNA libraries were purified after each of the enzymatic reactions using the AMPure XP beads (Beckman Coulter, A63882). All the samples were loaded on a SpotON flow cell (Oxford Nanopore Technologies, R9.4.1) and sequenced on a MinION (Oxford Nanopore Technologies, Mk1C).