title: GSM7163033: CCB1244 (pXsa-t15-r1); Bacillus subtilis; RNA-Seq
library_construction_protocol: Total RNA was isolated from 10 mL culture (pelleted and frozen) by the glass beads/phenol method described previously (Bechhofer DH et al. 2008, Methods Enzymol 447:259–276.). RNA samples were treated with RQ DNase Promega (37°C for 20 min) to remove potential contaminating chromosomal DNA. Ribosomal RNA was removed using the RiboZero kit (Illumina), and 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 the manufacturer's instructions. cDNA concentration and quality were checked by Bioanalyser (Agilent). The 6 samples were normalised to 2 nM, multiplexed and denatured at a concentration of 1 nM using 0.1 N NaOH (5 ﻿min at room temperature) before dilution to 10 pM and loading on a HiSeq Rapid SE65