GSM9578179: P. aeruginosa PAO1, 18 °C, 2 days, biol rep 2; Pseudomonas aeruginosa PAO1; RNA-Seq
To mechanically assist cell lysis, the samples were transferred to ZR BashingBead Lysis Tubes (0.1 mm and 0.5 mm, Zymo Research Europe, Freiburg, Germany) and homogenized for 10 min (Vortex-Genie 2, SI-0236, Scientific Industries, Inc., New York, USA). The samples were centrifuged at 10,000 × g at room temperature for 1 min and the supernatant was transferred to RNase-free tubes (AM12400, Thermo Fisher Scientific, Waltham, MA, USA). RNA was isolated and purified using the Direct-zol RNA Miniprep Kit (R2050, Zymo Research Europe, Freiburg, Germany) according to the manufacturer's instructions with subsequent quality and concentration check by the NanoPhotometer® NP80 (Implen, Munich, Germany). RNA was stored at -80 °C. Library preparation, sequencing, and data analysis were performed by Novogene GmbH (Munich, Germany), ribosomal RNA was eliminated from the total RNA, accompanied by ethanol precipitation. For quantification of the library, Qubit and real-time PCR were used, and the Agilent 2100 Bioanalyzer for size distribution detection. The libraries were pooled according to their effective concentration and the required data volume and sequenced on an Illumina NovaSeq 6000 platform.