RNA-Seq of Alcanivorax Borkumensis growing on different concentrations of iron
RNA was extracted from 2 ml of A.borkumensis SK2 pure cultures, at mid-log phase of growth and under described conditions, using a Masterpure Complete DNA and RNA purification RNA kit (Epicentre). The cells were harvested by centrifugation at 30005000 g at 4C. Extraction was carried out according to the manufacturer's instructions. RNA was stored in isopropanol at 20C before precipitation. RNA was resuspended in 50 l of RNase-free water. Extracted RNA was treated with a TURBO DNA-free kit (Ambion) to eliminate any residual DNA from the final elution. RNA samples quality and concentration were determined using Qubit 3.0 fluorometer (Thermo Fisher Scientific, Italy). The integrity and quality of RNA samples were assessed using the Agilent 2100 Bioanalyzer from Agilent Technologies. Following rRNA depletion and DNAse treatment, sequencing was conducted by FISABIO in Valencia, Spain. using the Illumina NextSeq Mid Output platform with 2100bp short insert single reads libraries (NextSeq Reagent Kit v2.5).