GSM5736067: NAC 8000 mg/L 30 min R1; Pseudomonas aeruginosa; RNA-Seq
Total RNA extraction was performed using the SV Total RNA Isolation System (Promega, Madison, WI, USA) with minor modifications. Briefly, 1 mL of each culture was transferred to sterile tubes and cells harvested by centrifugation (12,000 xg for 10 minutes) at 5 °C. The cell pellet was resuspended in 100 µL of 0.4 mg/mL lysozyme in TE buffer and incubated at room temperature for 5 minutes. RNA was purified according to the manufacturer's RNA purification by centrifugation protocol, including a second elution step performed with 50 μL of nuclease-free water. The purified RNA was then stored at –70 °C prior to ribosomal RNA depletion and sequencing. Total RNA quantity and quality, ribosomal RNA (rRNA) depletion, cDNA library construction and Illumina HiSeq 4000 platform-based RNA-sequencing were performed by Eurofins Genomics Europe Sequencing (Constance, Germany). The transcriptome libraries were strand-specific and single-end sequenced with 50-bp reads, for a total of 10 million reads per sample. At least 20 ng of rRNA-depleted RNA was used as input.