RNA-Seq of wildtype R20291 NO donor treated
RNA library preparation was performed using the Illumina Stranded Total RNA Prep kit, with rRNA depletion done through the Ribo-Zero Plus Microbiome kit. The rRNA-depleted RNA samples were fragmented into a size range of 200300 bases and converted to double-stranded cDNA. The cDNA was end-repaired and adenylated at the 3' end to enable the ligation of Illumina adapters with an overhang. Each library fragment was PCR-amplified with a unique barcoded adapter to allow for multiplex sequencing. Libraries were sequenced on the NovaSeq X system using a 10B flow cell to generate 150-base paired-end reads.