title: PAO1_Acetate_2
organism: Pseudomonas aeruginosa PAO1
description: Protocols: P. aeruginosa strain PAO1 was grown in 40 ml MOPS with acetate or glycerol as sole carbon sources (quadruplicate), 37 °C with shaking (250 rpm) in baffled flasks (500 ml). 5 ml of culture was harvested from each sample at OD600 of 0.5 (exponential growth) and added to an equal volume of RNAlater® RNA stabilization solution. Samples were then centrifuged at 4000 rpm for 15 min (4 °C) and the pellets were stored at - 80 °C. 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). Eight 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: 2019-10-05T04:02:54Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-09-25T15:04:34Z
attributes.tag: External Id
attributes.value: SAMEA5972742
attributes.tag: INSDC center name
attributes.value: UNIVERSITY OF CAMBRIDGE
attributes.tag: INSDC first public
attributes.value: 2019-10-05T04:02:54Z
attributes.tag: INSDC last update
attributes.value: 2019-09-25T15:04:34Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-8374:PAO1_Acetate_2
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: growth condition
attributes.value: MOPS Acetate
attributes.tag: organism part
attributes.value: whole organism
attributes.tag: sample name
attributes.value: E-MTAB-8374:PAO1_Acetate_2
attributes.tag: scientific_name
attributes.value: Pseudomonas aeruginosa PAO1