GSM7033647: PA_in_BC_SN_FeHigh_2; Pseudomonas aeruginosa PAO1; RNA-Seq
The cultures were transferred onto ice. We then distributed them to 50 ml tubes (Greiner) containing 5 ml ice cold stop solution (10% phenol buffered with Tris-HCl to pH8 (Sigma-Aldrich, Switzerland) and 90% ethanol). The suspension was mixed by manual inversion of tubes and centrifuged at 4°C, for 5 minutes at 9000 rpm. The supernatant was discarded, while the cell pellet was flash-frozen in liquid nitrogen and stored at -80°C. RNA was harvested using hot phenol protocol (Pessi et al 2007). Afterwards cDNA was produced. The supernatant treatments were each mixed with CAA and independently generated supernatants. So we used three biological repliactes of each supernatant to generate three independend biological replicates of our PAO1 culture for RNA isolation. RNA libraries were prepared using Encore Complete Prokaryotic RNA-Seq DR Muliplex System