title: RNA-seq of Streptomyces coelicolor hupA and hupS deletion strain
design: RNA-seq of Streptomyces coelicolor hupA and hupS deletion strain
library_construction_protocol: For RNA-seq, total RNA was isolated from 30 ml YEME/TSB cultures. Cultures were inoculated with S. coelicolor and cultivated in flasks with spring coils for 24-36h in 30°C. For osmotic stress experiment growth medium was supplemented with 0.5M NaCl. Mycelia were collected at two time points (exponential and early stationary growth, determined individually for each strain based on the growth curve (Fig. S3)) by collecting 2 ml from the culture and centrifugation. Cell pellets were frozen and stored at -70°C for subsequent RNA isolation.  RNA was isolated using the RNeasy Mini Kit (Qiagen) following manufacturer's instructions, DNA digestions was performed using on column digestion with DNase-I (Qiagen) and TURBO DNase I (Ambion). RNA quality and concentration was measured using Nanodrop (Thermo Fisher Scientific) and Qubit (Thermo Fisher Scientific).  The absence of DNA in the sample was confirmed using PCR. Library preparation was performed by Genewiz (Germany).