GSM7106012
Cultures were then treated with RNAprotect (Qiagen) for 5 min at RT to stabilize mRNA, and then collected by centrifugation (6,000 X g, 5 min). Total RNA was isolated by hot-phenol extraction, and further purified using the RNeasy Kit (Qiagen) with on-column DNase I treatment. The eluates were treated with Turbo DNase (Invitrogen) for 30 min before a second round of purification using the RNeasy Kit.  rRNA was depleted using the NEBNext® rRNA Depletion kit (Bacteria) and purified from the enzymatic reactions using RNAClean XP magnetic beads (Beckman Coulter) following manufacturer's instructions. RNA quality and ribosomal depletion was assessed using BioAnalyzer (Agilent). RNA-seq libraries were made and sequenced by the Genomics Core Facility at Penn State University.