title: GSM1616445: d6977_A; Pseudomonas aeruginosa; RNA-Seq
library_construction_protocol: Total RNA was extracted using the RNEasy kit (Qiagen) according to the manufacturer's instructions with minor modifications. Briefly, frozen pellets were resuspeded in 215 ul TE buffer containing 15 mg/ml lysozyme (Sigma) and 1.4 mg/ml proteinase K (Qiagen). Samples were incubated with shaking for 8 minutes at 37 degrees C to disrupt the cell walls.  700 ul buffer RLT and 500 ul 100% ethanol were added and the mixture was applied to the Qiagen column. On-column DNA digestion, washing, and elution were carried out according to instructions. 10 ug total RNA was then further treated with the Turbo DNase free kit (Ambion) according to instructions. 3.8 ug total RNA was subjected to ribosomal RNA depletion using the Ribo-Zero Magnetic Kit for gram negative bacteria (Epicentre). Libraries for RNA Seq were constructed from the rRNA-depleted samples using the NEBNext mRNA Library Prep Master Mix Set for Illumina, according to the manufacturer's instructions.
identifiers.namespace: GEO
identifiers.id: GSM1616445