E. coli MG1655 pJV300 stationary phase

Over-night cultures grown from three single colonies were diluted 1:100 in LB and grown with shaking at 37 ?C for 6h. Bacteria were pelleted by centrifugation and fast-freezed in the presence of 0.9 mg/ml lysosyme in TE. Frozen cells were thawed and re-frozen twice in 37 ?C and liquid nitrogen, respectively. Total RNA was extracted using TRI-reagent and used for construction of RNA-seq libraries according to the RNAtag-seq method. Libraries were paired-end sequenced using Illumina NextSeq 500 platform.