GSM4224634: WT M9/rdm/glu, RNA; Escherichia coli; RNA-Seq
Samples for RNA isolation were preserved immediately prior to rifampicin addition by dilution in a 5x excess of DNA/RNA Shield (Zymo); the RNA samples were then stored at -80o C until purification. RNA was isolated using a Zymo QuickRNA microprep kit following the manufacturer's instructions, including the on-column DNase digestion. Purified RNA was quantified using RiboGreen (Invitrogen), and then ribosome-depleted using the Illumina RiboZero Gram-negative bacteria kit according to the manufacturer's instructions, with the input RNA amount and all reaction volumes cut in half. Final recovery of the ribo-depleted RNA was accomplished using the modified Zymo spin column protocol present in the RiboZero documentation. Epicentre ScriptSeq v2 prep kit