Transcriptome sequencing of Riemerella anatipestifer CH-1 and CH-1_zntR under TSB condition

Identifying the differentially expressed genes of Riemerella anatipestifer CH-1 and CH-1_zntR under normal condition through RNA sequencing. For RNA extraction, the WT and ΔzntR strains were cultured at 37°C in TSB to an OD600 of ~1.5, and adjusted to OD600 = 0.5 using PBS. Total RNA was extracted from the bacterial pellet using an RNAprep Pure Cell/Bacteria Kit (TIANGEN). We used a NanoDrop 2000 to assess the concentration and quality of RNA samples and then sent samples to Shanghai Majorbio Bio-Pharm Technology Co., Ltd. for RNA sequencing (RNA-seq). Library preparation for RNA transcriptome sequencing was performed using the TruSeqTM Stranded Total RNA Library Prep Kit. The rRNA was first removed from total RNA, and the mRNA was cleaved into ~200 bp fragments using fragmentation buffer. Single-stranded cDNA was synthesized with random primers and the second-strand cDNA was synthesized with dUTP instead of dTTP. The ends of double-stranded cDNA were blunted with the End Repair Mix and an A base was added at the 3’ end. The cDNA was then digested with the UNG enzyme to remove the second-strand. Finally, the remaining first-strand cDNA was quantified using Qubit 4.0 and sequenced on the NovaSeq XPlus platform