title: GSM8714658: RNase J mutant vegetative, 12h, biol rep 1; Streptomyces venezuelae; RNA-Seq
library_construction_protocol: Cells were lysed by vortexing pellets for 2 min with an approximately equal volume of sterile glass beads and ice cold lysis solution (4M guanidine thiocyanate, 25 mM trisodium citrate, 0.5%v/v sodium N-lauroylsarcosinate, 0.8%v/v β-mercaptoethanol). An equal volume of phenol:chloroform:isoamyl alcohol (50:50:1, pH 4.6) was added to the lysate and samples were vortexed on and off in thirty second intervals to emulsify the solutions. Samples were then centrifuged at 7,800 ×g and the aqueous phase containing the RNA was recovered. Total RNA was precipitated by adding an equal volume of ice-cold isopropanol (100%), 0.1 volume of sodium acetate buffer (3 M, pH 6), and 2 μL of RNA grade glycogen (ThermoFisher Scientific).The total RNA sample was DNase treated post-extraction using TURBO DNase buffer and enzyme (Fisher Scientific). Total RNA was extracted from the reaction mixture with phenol:chloroform:isoamyl alcohol (50:50:1, pH=4.6). RNA-sequencing was conducted by the MOBIX facility at McMaster University using paired-end technology on the Illumina MiSeq v3 platform.