GSM5510109: Reactor 1 under Anoxic conditions; Paracoccus denitrificans; RNA-Seq
QIAGEN RNeasy Mini Kit was used for purification of total RNA from bacteria. Bacterial cells were disrupted and homogenized by bead-milling in a guanidine-thiocyanate–containing lysis buffer. After addition of ethanol, the sample was loaded onto an RNeasy Mini spin column. Total RNA binds to the RNeasy silica-membrane, contaminants were efficiently washed away, and high-quality RNA was eluted in RNase-free water. For Total RNA sequencing service, RNA integrity was checked with Agilent Technologies 2100 Bioanalyzer. Ribosomal RNA was removed, followed by fragmentation with divalent cation buffers in elevated temperature. The sequencing library was prepared following Illumina's TruSeq-stranded-total-RNA-sample preparation protocol. Quality control analysis and quantification of the sequencing library were performed using Agilent Technologies 2100 Bioanalyzer High Sensitivity DNA Chip. Paired-ended sequencing was performed on Illumina's NovaSeq 6000 sequencing system.