Illumina HiSeq 2000 sequencing; Pseudmonas aeruginosa PAO1 wild-type cultured in MOPS Glycerol compared to MOPS Glycerol Hypoxia (restricted oxygen).
Pseudmonas aeruginosa PAO1 wild-type cultured in MOPS Glycerol compared to MOPS Glycerol Hypoxia (restricted oxygen).
Pseudmonas aeruginosa PAO1 wild-type cultured in MOPS Glycerol compared to MOPS Glycerol Hypoxia (restricted oxygen). P. aeruginosa strain PAO1 was grown in 40 ml MOPS with glycerol as sole carbon source (triplicate), 37 °C with shaking (250 rpm) in baffled flasks (500 ml). For the restricted oxygen condition, P. aerugionosa was cultured in the same conditions, except non-baffled flasks were used, and the shaking speed was 80 rpm (gentle aggitation). A thin layer 10 ml of mineral oil was overlaid on top of these cultures to restrict oxygen transfer. Total RNA was isolated as described in [90], followed by phenol-chloroform-isoamyl alcohol extraction and ethanol precipitation. Purity and concentration of the DNase treated (Turbo DNA-Free, Applied Biosystems) RNA was estimated with a NanoDrop ND-1000 spectrophotometer (NanoDrop technologies Inc, USA) and using agarose gel electrophoresis. Ribosomal RNA (rRNA) was then depleted from each sample (5 µg each) using the bacterial Ribo-Zero rRNA Removal Kit (Illumina). The integrity of the RNA was evaluated using an RNA 6000 Nano LabChip and an Agilent 2100 Bioanalyzer (Agilent Technologies, Germany).  Six indexed, strand-specific cDNA libraries were prepared, and samples were sequenced on an Illumina HiSeq 2000 with a 51-bp single-end read length (GATC Biotech, Germany).
Experimental Factor: environmental stress
hypoxia