RNA-Seq of Streptomyces venezuelae: 24h after mock treatment (water)
Streptomyces venezuelae NRRL B-65442 was grown in MYM medium, containing 2.1 g/L MOPS (Sigma). Ethanol shock was performed after 7 h of cultivation by adding 1.5 mL (6% v/v) of absolute ethanol or 1.5 mL distilled sterile water (control) to the cultures. The fermentations were stopped after 18, 24 and 48 h. All the fermentations have been performed in triplets. 1.5 mL samples were immediately taken from each culture and transferred in sterile RNAse-free tubes containing 3 mL RNAprotect Bacteria Reagent (QIAGEN, Germany). The samples were vortexed for 10 sec, rested on table for 5 min, transferred into 2.5 mL sterile RNAse-free Eppendorf tubes and centrifuged for 12 min at 4000 rpm, 4 C. Supernatants were removed and cells frozen immediately at -80 C. One tube with the cells per sample was taken for RNAseq analysis.