Illumina MiSeq paired end sequencing; Global gene transcription analysis in Streptomyces coelicolor strains with modification of satKR genes.
RNA was isolated from vegetative hyphae of S. coelicolor cultivated in 30 mL liquid YEME/TSB medium for 18 hours. Mycelia were collected by centrifugation, frozen and stored at -70°C for subsequent RNA isolation. medium: liquid YEME/TSB; 30 mL, 18 hours of growth, 30C, 180 rpm. RNA was isolated using the procedure described previously by Moody et al. (Moody et al., 2013), digested with TURBO DNase I (Invitrogen, USA). RNA libraries were prepared for sequencing using standard Illumina protocols (NEBNext Ultra II Directional RNA Library Prep Kit). rRNA was removed using Ribo-Zero Ribosomal RNA Removal Kit (Illumina).