GSM6500810: PAO1161 PA3973 overproducer, biological rep1; Pseudomonas aeruginosa; RNA-Seq
2 ml of cultures which reached optical density 0.4-0.6 at 600 nm were mixed with 4 ml of RNAprotect Bacteria Reagent (Qiagen). RNA was isolated with an RNeasy mini-kit (Qiagen) according to the manufacturer's protocol. Total RNA was digested with DNase (TURBO DNA-free Kit, Ambion) to eliminate genomic DNA. rRNA was depleted using Ribo-Zero™ rRNA Removal Kit (Bacteria) (MRZMB126, Illumina) according to manufacturer instructions. Libraries were prepared according to instructions accompanying the NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina® (NEB, E7420). Libraries were sequenced using standard Illumina protocols.