GSM6454140: ST28, sigE null mutant ST28, 6 hours, biol rep 2; Mycobacterium tuberculosis; RNA-Seq
Total RNA was extracted from 35 mL cultures as previously described [10.1128/JB.00756-09]. Two or three DNase treatments were performed to remove DNA contamination from the samples. The DNA contamination was verified using 5ng of nucleic acids samples. RNA was quantified by spectrophotometer (Nanodrop) and its quality was verified by bioanalyzer (Agilent RNA 6000 Nano kit). Only samples with RIN values of 8/9 were used for the following steps. Before sequencing, the Ambion ERCC Spike-In Control Mixes from Thermo Fisher Scientific were added to the M. tuberculosis RNAs following the manufacturer instructions. Tagged libraries were prepared with the Illumina TruSeq stranded protocol with depletion of ribosomal RNA (rRNA), pooled and subjected to 2x100 bp paired-end sequencing.