RNA sequencing of Escherichia coli single mutants and ancestral genotype at high temperature
Total RNA was extracted using Qiagen RNeasy kits. RNA concentrations were measured with Qubit RNA HS assay kits and RNA quality was assessed by running an Agilent RNA-Nano chip on a bioanalyzer. We enriched for mRNA by the removal of rRNA using NEBNext rRNA depletion kits for bacteria. We prepared the RNA for Illumina sequencing using the NEB Ultra II Directional RNA Library Prep kit. All samples were uniquely barcoded and multiplexed for sequencing with Illumina NovaSeq at the UCI Genomics High Throughput Facility.