GSM4826695: sg-CTRL r3; Pseudomonas aeruginosa; RNA-Seq
bacterial pellets were resuspended in 1ml Tri Reagent (Sigma Aldrich T9424) and incubated for 5 min. 200 ul Chloroform were added, vortexed and incubated for 5 min. Samples were centrifuged for 10 min at 4°C and the aqueous phase was mixed with 200 ul 100% EtOH and transferred to PureLink RNAMini Columns (Invitrogen). Subsequent washing and elution was performed following manufactorer's instructions. Contaminating DNA was removed using DNA-free kit (Apllied Bioystems). RNA was stored at -80 °C. Libraries were constructed using NuGEN Ovation Complete Prokaryotic RNA-Seq DR Multiplex Systems kit. Preparation was performed following manufactorer's instructions