title: GSM4491547: WT rep3; Mycolicibacterium smegmatis; RNA-Seq
library_construction_protocol: Bacterial cultures were grown up to the logarithmic phase (OD ~ 1.0), centrifuged (4°C, 3000 rpm) and total RNA was isolated by phenol-chloroform extraction after cell disruption by Bead Beater with 0.1 mm zirconia beads (BioSpec Products, USA) as previously described (Rustad et al., 2009). After isolation, RNA was treated with Turbo DNase (Life Technologies, USA) to remove traces of genomic DNA, and purified with the RNeasy mini kit (Qiagen, Netherlands). Amounts and purity of RNA were determined spectrophotometrically; integrity of RNA was assessed in 1% agarose gel. RNA samples were depleted of rRNA using Ribo-Zero rRNA Removal Kit (Bacteria). Sequencing libraries  were generated using the resulting ribosomal transcript-depleted RNA and NEBNext Ultra II Directional  RNA Library Prep Kit (NEB, USA) according to the manufacturers' protocol. Sequencing was performed in  triplicates using the Illumina HiSeq2500 as the pair-ended 150 nt-long reads.
attributes.tag: GEO Accession
attributes.value: GSM4491547