RNA-seq of Aeromonas hydrophila
Total RNA was extracted using Trizol reagent (Invitrogen Life Technologies). The quality and integrity of the RNA were assessed with a NanoDrop spectrophotometer (Thermo Scientific) and a Bioanalyzer 2100 system (Agilent Technologies). The Zymo-Seq RiboFree Total RNA Library Kit was used to deplete rRNA from the total RNA sample. First-strand cDNA synthesis was performed using random oligonucleotides and SuperScript III reverse transcriptase. Subsequently, RNaseH is used to degrade the RNA strand. In the DNA polymerase I system, dNTP and dUTP are used instead of dTTP as raw materials to synthesize the second strand of cDNA. Exonuclease and polymerase activities were employed to convert the remaining overhangs into blunt ends, followed by enzyme removal.