alias: GSM2464565
title: GSM2464565: A CCB604 (W168 [delta]yacP + pDG-yacP) - IPTG; Bacillus subtilis; RNA-Seq
library_construction_protocol: Total RNA was isolated from 20 mL culture (pelleted and frozen) by the glass beads/phenol method described previously (Bechhofer et al, 2008). RNA samples were treated with RQ DNase Promega (37°C for 20 minutes) to remove potential contaminating chromosomal DNA. Ribosomal RNA was removed from 5 mg total RNA using RiboZero kit from Epicenter, according to the manufacturer's instructions. Ribosomal RNA depletion and overall RNA quality was analysed by Bioanalyser (Agilent). cDNA libraries were prepared using the Smarter Stranded RNA-Seq Kit (Clontech) with adapters for multiplexing, according to manufacturer's instructions. RNA concentration and quality was checked by Bioanalyser (Agilent). The 12 samples were normalized to 2nM, multiplexed and denatured at a concentration of 1nM using 0.1N NaOH (5 minutes at room temperature) before dilution to 10pM and loading on a HiSeq2000 flowcell.
identifiers.namespace: GEO
identifiers.id: GSM2464565