title: Halobacterium salinarum NRC-1; wild type; pMTF overexpressing cMyc tag; late-exponential phase; biological replicate 2; technical replicate 1
design: Halobacterium salinarum NRC-1 carrying the pMTF vector overexpressing the 13-cMyc tag was cultured in mevinolin-supplemented (20 ug/mL) complex medium (CM) until late-exponential phase (OD600nm = 0.75). The cultures were grown at 37C, under light exposure, and constant agitation (125 RPM). Samples of 20 mL were processed according to the protein-RNA co-immunoprecipitation protocol. Please, check our manuscript for details. We submitted the co-immunoprecipitated samples to protein digestion using proteinase K (Fermentas). Then, we purified the RNA using the MinElute Reaction Cleanup Kit (QIAGEN) with a DNase treatment step. The RNA samples were quantified by the Quant-iT RiboGreen RNA Assay (Invitrogen) and prepared for sequencing using the TruSeq mRNA Stranded kit (Illumina). Before sequencing, to equalize the concentrations, another round of quantification was performed by using the KAPA Library Quant kit (KapaBiosystems). Samples were sequenced using the MiSeq Reagent v2 kit (Illumina; 50 cycles; single-end mode) in a MiSeq (Illumina) instrument. We sequenced one technical replicate for each one of the two biological replicates. This is the technical replicate 1 of the biological replicate 2. The following sequence may be used to trim adapters: AGATCGGAAGAGCACACGTCTGAACTCCAGTCAC.