GSM5009682: G. diazotrophicus cells, 0 µM FeCl3 R1; Gluconacetobacter diazotrophicus; RNA-Seq
Total RNA was isolated with Trizol in accordance with the manufacturer's protocol (Life Technologies) and treated with DNase I (Epicenter) in order to remove genomic DNA contamination. RNA purity was quantified using Qubit (Life Technologies). Seven micrograms of total RNA were used to ribosomal RNA (rRNA) depletion with MICROBExpressTM kit. The efficiency of depletion was evaluated in agarose gel electrophoresis (1%) followed by quantification of the total RNA with Agilent 2100 Bioanalyzer (Agilent). A total of 500 ng mRNA was used for the contruction of a sequencing library using the standard protocol of the SOLid Total RNAseq Kit (Life Technologies). The libraries were barcoded by using the SOLiD Transcriptome Multiplexing Kit (Life Technologies). The emulsion PCR and sequencing were performed according to Ion One TouchTM 200 Template Kit v2 DL and Ion PITM Sequencing 200 Kit v2 using the standard Life Technologies protocols, respectively. The Ion Proton Semiconductor Sequence (Life) was used to sequence twelve libraries generated from three biological replicates from each independent treatment.