title: NextSeq 500 paired end sequencing; RNA-seq of Pseudomonas aeruginosa PAO1 wild-type and ersA (sRNA) knock-out mutant
design: RNA-seq of Pseudomonas aeruginosa PAO1 wild-type and ersA (sRNA) knock-out mutant
library_construction_protocol: Bacteria were grown in BHI medium at 37°C under shaking conditions (200 rpm). Cells were harvested by centrifugation after reaching OD600 2.7 . Pellets were frozen at -80°C. Bacterial pellets were resupended in 200 µl of 1x Tris-EDTA buffer (pH 8.0) containing 100 µg/ml lysozyme and 20 µg/ml Proteinase K. RNA was extracted using Trizol reagent according the provided protocol. RNA was purified from aqueous phase using Zymo Research RNA Clean & concentrator kit. rRNA was depleted using Ribo-Zero rRNA Removal Kit (Gram-Negative Bacteria) and library was prepared following protocol provided with Illumina TruSeq Stranded mRNA Library Prep Kit.
attributes.tag: Experimental Factor: genotype
attributes.value: wild type genotype