L7
RNA Extraction and Preparation of Sequencing Libraries for RNA-Seq. RNA was extracted using RNeasy(r) Mini (Qiagen, cat# 74106), with a mild modification. Bacterial pellets were lysed using PowerBead Pro tubes (Qiagen, cat#19301) with RLT buffer, on a horizontal vortex adapter for 10 minutes. The beads were centrifuged, and the resulting supernatant was cleaned following manufacturer's instructions. RNA was assessed using Tapestation (Agilent), with RIN scores of 8.5 and above. Ribosomal RNA was depleted using Ribo-zero Plus (Illumina, cat#20037135), and sequencing libraires were prepared using NEBNext Ultra II Directional RNA Library Prep Kit (NEB, cat# E7760L). Paired end 150bp sequencing was carried out on an Illumina NovaSeq 6000, with 30 million reads per sample.