GSM6044833: RhlR biol rep 3; Pseudomonas aeruginosa; RNA-Seq
mRNA was preserved immediately upon cell harvest using Qiagen RNAprotect. Cells were lysed using sonication and RNA was harvested using RNeasy Mini Kit (Qiagen) including the on-column DNase I digestion step. In addition, eluted RNA was treated for 1 h at 37°C with TURBO DNase (0.2 U per μg of RNA; Ambion) and with SUPERase-In (0.4 U per μg of RNA; Ambion). DNase I was removed with the RNeasy Column Purification Kit (Qiagen). 3 µg of total RNA was used for the construction of sequencing libraries. rRNA depletion was performed using NEBNext rRNA Depletion Kit (Bacteria). RNA sequencing library preparation was performed using NEBNext Ultra II RNA Library Prep Kit for Illumina by following the manufacturer's recommendations (NEB, Ipswich, MA, USA).