Identifications of transcription start sites (TSSs) in the extremophyle archeon H. salinarum NRC-1
Total RNAs were treated with Turbo DNase (Ambion) and incubated with Terminator 5'-phosphate-dependent exonuclease (Epicentre) (TEX+ sample) or only in buffer reaction (TEX- sample) at 30ºC for 60 min, at proportion of 1 U TEX per 1 µg total RNA. Reaction products were purified with RNeasy MinElute Cleanup Kit (QIAGEN) and incubated with 1 U of Tobacco Acid Pyrophosphatase (TAP) (Epicentre) at 37ºC for 1 hour in order to generate 5'-mono-phosphates RNAs able to bind to sequencing adapters. Reactions were purified again with RNeasy MinElute Cleanup Kit (QIAGEN).