title: GSM5745214: 3610, control 2; Bacillus subtilis subsp. subtilis NCIB 3610 = ATCC 6051 = DSM 10; RNA-Seq
library_construction_protocol: Liquid growth cultures of B. subtilis were grown overnight in Luria–Bertani (LB) medium at 30°C. Overnight cultures were diluted and, after 3-4 hours (exponential phase), B. subtilis cells were treated with Tse1 or with control buffer. Cultures were grown for 3 h.  Cells were harvested by centrifugation at 8000 g for 10 min and suspended immediately in stop solution (95% (v/v) ethanol, 5% (v/v) phenol) and pelleted by centrifugation. After that, total RNA was extracted with Trizol (Invitrogen). Removal of DNA was carried out by treatment with Nucleo-Spin RNA Plant (Macherey–Nagel) kit. The integrity and quality of total RNA was assessed with an Agilent 2100 Bioanalyzer (Agilent Technologies). RNA libraries were prepared for sequencing using standard Illumina protocols