GSM9177267: Control (PBS) replicate 1; Escherichia coli; RNA-Seq
Bacterial were collected by centrifugation at 4000×g at 4 °C for 12 min. Total RNA was extracted using TRIzol reagent (Thermo Fisher, USA) according to the manufacturer's instructions. RNA integrity was assessed using agarose gel electrophoresis and an Agilent 2100 system. mRNA was isolated by removing rRNA, fragmented to ~200 bp, and used for first-strand cDNA synthesis with random primers. Second-strand cDNA synthesis was performed with dUTP substitution for dTTP. Libraries were prepared using a TruSeq Stranded Total RNA Library Prep Kit (Illumina, USA), and the second strand was digested with UNG before PCR amplification.