GSM9083428: 43_T0_3; Pseudomonas aeruginosa PAO1; RNA-Seq
90 mg/mL Lysozyme (Roche), 10 mL Proteinase K (Qiagen), and 1 mL warm Tri-reagent 37°C (Sigma) were added to pelleted cells. After 5 min of incubation at 65°C, 200 mL chloroform was added. The solution was centrifuged for 15 min at 20,817 g, and the upper liquid phase was transferred into 80% ethanol. The RNA was then extracted using an RNeasy minikit (Qiagen) according to the manufacturer's protocol. The library was constructed with NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina (NEB) according to the manufacturer's instructions using 30 ng of depleted RNA.