title: Gly_Hypoxia_3
organism: Pseudomonas aeruginosa PAO1
description: Protocols: 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).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2019-12-02T04:02:53Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-10-16T14:55:55Z
attributes.tag: External Id
attributes.value: SAMEA6087122
attributes.tag: INSDC center name
attributes.value: UNIVERSITY OF CAMBRIDGE
attributes.tag: INSDC first public
attributes.value: 2019-12-02T04:02:53Z
attributes.tag: INSDC last update
attributes.value: 2019-10-16T14:55:55Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-8442:Gly_Hypoxia_3
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: growth condition
attributes.value: MOPS Glycerol
attributes.tag: organism part
attributes.value: whole organism
attributes.tag: sample name
attributes.value: E-MTAB-8442:Gly_Hypoxia_3
attributes.tag: scientific_name
attributes.value: Pseudomonas aeruginosa PAO1