title: GSM8559668: Haloferax volcanii cells, rosR deletion strain, rep3; Haloferax volcanii DS2; RNA-Seq
library_construction_protocol: RNA was extracted using an Absolutely RNA Miniprep kit according to manufacturer instructions (Agilent, Santa Clara, CA). RNA quality was measured using RNA 6000 Nano kit on a 2100 Bioanalyzer (Aglient, Santa Clara, CA). PCR to verify lack of DNA contamination was  conducted on 300 ng of RNA sample using species-specific primers listed in Supplementary Table S2. Ribosomal RNA depletion was performed from 300 ng of each sample using NEBNextR RNA Depletion Core Reagent (#E7870 New England Biolabs) according to the manufacturer instructions. The resultant mRNA was purified using NEBNext RNA sample purification beads and eluted in 7 l of nuclease-free water. 1l of each sample was used for quantification and quality verification (free of rRNA) in a second round of Agilent Bioanalyzer analysis using 2100 Prokaryote Total RNA Nano kit (Aglient, Santa Clara, CA). Libraries were constructed using 5 ml of mRNA using the Ultra II Directional RNA Library Preparation Kit (Agilent, Santa Clara. CA). Indexes were designed according to the kit specifications and obtained from Integrated DNA Technologies. Resultant libraries were tested for quality and quantity by Bioanalyzer using the 2100 expert High Sensitivity DNA Assay chip (Agilent Technologies)