title: GSM9491799: Salmonella Typhimurium Δ5msr, mid log, biol rep 3; Salmonella enterica subsp. enterica serovar Typhimurium; RNA-Seq
library_construction_protocol: Frozen bacterial pellets from 10 mL cultures were subjected to RNA extraction using the Qiagen RNeasy Mini Kit according to the manufacturer's instructions. RNA was quantified using a Qubit fluorometer, and RNA integrity was assessed using High Sensitivity RNA ScreenTape to determine RIN values. A total of 1.2 µg of high-quality RNA was subjected to rRNA depletion using the NEBNext® rRNA Depletion Kit (Bacteria) following the manufacturer's protocol. Double stranded cDNA libraries were prepared using the NEBNext® Ultra II Directional RNA Library Prep Kit for Illumina. Library construction involved RNA fragmentation, first-strand cDNA synthesis using random primers, second-strand cDNA synthesis, end repair, adapter ligation, removal of excess adapters using purification beads, PCR enrichment, and cleanup of PCR products. Final libraries were quantified using a Qubit fluorometer and assessed for fragment size distribution using High Sensitivity D1000 ScreenTape.