Bulk RNA-sequencing of USA300 LAC grown with or without itaconate
LAC was grown in LB with or without 30 mM itaconate to late exponential phase. Bacterial pellets were incubated in a cell wall lysis mixture at 37 C for 45 min. TRK lysis buffer and 70% ethanol were added, and samples were transferred to E.Z.N.A RNA isolation columns. RNA was isolated following the manufacturers instructions and treated with DNase using the DNAfree DNA removal kit . The RNA was precipitated with 0.1 volume 3 M sodium acetate and 3 volumes of 100% ethanol, recovered by centrifugation and washed with ice cold 70% ethanol. A ribosomal RNA-depleted cDNA library was prepared according to the manufacturers instructions using the Universal Prokaryotic RNA-Seq Prokaryotic AnyDeplete kit.