GSM3418164: Hospital-acquired strain; Pseudomonas aeruginosa; RNA-Seq
Bacteria were lysed and homogenized using TRIzol reagent (Thermo Fisher Scientific). The homogenates were separated into aqueous, interphase, and organic layers after chloroform was added. RNA was precipitated from the aqueous layer using isopropanol. The precipitated RNA was washed with cold 75% ethanol and then solubilized using RNase-free water. The contaminated DNA was removed after DNase I (Takara) treatment. RNA samples were reverse-transcribed into cDNA using Ovation Complete Prokaryotic RNA-Seq DR Multiplex System 9-16 (NuGEN), and cDNA fragmented using S220 system (Covaris) then constructed fragment cDNA Library.