title: GSM9503135: AB5075_in_vivo_1; Acinetobacter baumannii; RNA-Seq
library_construction_protocol: Total RNA was extracted using Total RNA Kit I (Omega) according to the manufacturer's instructions, with additional on column DNase I (Takara) treatment to remove genomic DNA. RNA quality and concentration were assessed using a Bioanalyzer (Agilent Technologies). Total RNA was extracted from the samples and treated with DNase I to digest genomic DNA. Eukaryotic mRNA was then enriched using magnetic beads conjugated with Oligo(dT); for prokaryotic samples, rRNA was removed with a commercial kit prior to the subsequent steps. mRNA fragmentation reagent was added to shear the mRNA into short fragments at an optimal temperature in a Thermomixer. The sheared mRNA was used as the template to synthesize first-strand cDNA, followed by the preparation of a second-strand synthesis reaction system for second-strand cDNA generation. The resulting cDNA was purified and recovered with a kit, subjected to end repair, dA-tailing at the 3' ends, and adapter ligation, prior to fragment size selection and final PCR amplification. After the constructed libraries passed quality control using an Agilent 2100 Bioanalyzer and an ABI StepOnePlus Real-Time PCR System, high-throughput sequencing was performed on an Illumina NovaSeq 6000 platform.