title: GSM2109000: T0h_2h; Staphylococcus epidermidis; RNA-Seq
library_construction_protocol: BRC total RNA was extracted using RNeasy mini kit (QIAGEN) with minor alterations as described in França et al., 2012, PLoS One 7 (5): e37480. In brief, BRC were suspended in 500 µL of in the lyses buffer provided by the kit plus 500 µL of phenol. This supension was transfered into BeadBug tubes (1.0 mm Zirconium beads, triple-pure, high impact, Sigma) and using a MPBio 24 (MPBio) the cells were lysed (3 cyles at 6.5 meter/second for 35 seconds.Between each cycle samples were placed on ice for 5 minutes). Finally, the tubes were centrifuged and the suspension transfered into a new tube. An equal volume of 70 % of ethanol was added. The suspension was transfered in to the RNeasy mini kit columns and the manufacturer's instructions were strickly followed. Total RNA, eluted in 50 µL, was treated with TURBO DNase (Ambion) and 1:5 Acid-phenol:chloroform in order to degrade and isolate, respectively, genomic DNA. Additionally, total RNA was treated with MICROBEnrichTM (Ambion) and Ribo-ZeroTM rRNA removal kit for Gram-positive bacteria (Epicentre) in order to remove eukaryotic RNA and bacterial ribosomal RNA, respectively. Libraries were constructed using ScriptSeq RNAseq Library preparation kit (low input) and strictly following the manufacturer's indications.
attributes.tag: GEO Accession
attributes.value: GSM2109000