GSM6198659: wt untreated batch 1 rep2; Haloferax volcanii; RNA-Seq
The RNA was extracted using TRI Reagent® (#T9424, Sigma-Aldrich). To verify RNA integrity, the samples were separated by 0.8 % (w/v) agarose gel electrophoresis (at 90 V for 20 min) in 1X TAE buffer using 2X RNA Loading Dye (#R0641, Thermo Fisher Scientific). The samples were then treated with TURBO DNase-free kit (AM1907, Invitrogen) to remove traces of DNA. Only RNA samples with DNA below the limit of PCR detection were further processed. From each sample preparation, total RNA ≥ 6 μg in 50 μL with RIN value ≥ 6 was used to prepare a cDNA library and analyzed by high throughput sequencing (Novogene). The abundance of rRNA was reduced using the Ribo-ZeroTM Magnetic kit, and the cDNA was prepared using a 250 to 300 bp insert strand-specific library. The library was sequenced using the Illumina Platform PE150, Q30≥ 80% yielding 2G raw data / sample.