alias: PA_anoxic
title: Pseudomonas aeruginosa strain PA14 raw sequence reads (30 min anoxic)
design: The clinical isolate of Pseudomonas aeruginosa, PA14, was used in all  experiments.  The synthetic cystic fibrosis sputum medium was prepared a  described by Palmer et al. except that the concentration of FeSO4·7H2O was  increased to 100 µM and that of KNO3 to 100 mM.    Two biological replicates of each condition was prepared and analyzed in  this study. The following protocol describes the procedure for one single  sample.    For 30 min anoxic conditions, pre-cultures were incubated overnight in  SCFM at 37°C, shaking at 165 rpm. A 25 ml main culture was prepared from  the pre-culture in fresh SCFM with an OD600 nm of 0.05 and incubated at  37°C, shaking at 165 rpm, once the culture reaches OD600 nm of 0.4, and  shifted to anoxic chamber for 30 minutes.    Total RNA was isolated from cell pellets using the TRIzol reagent (Invitrogen)  according to the manufacturer’s instructions.  The samples were DNase I treated,  followed by phenol-chloroform-isoamyl alcohol extraction and ethanol  precipitation.   The MICROBExpress Kit (Ambion) was used to deplete rRNA from  total RNA samples.  Libraries were constructed using NEBNext® Ultra™ Directional  RNA Library Prep Kit from Illumina.  100 bp sequence reads were generated using  the Illumina HiSeq 2000 platform at the Vienna Biocenter Campus Science Support  Facility (http://www.csf.ac.at).
identifiers.namespace: University Vienna
identifiers.id: PA_anoxic