title: Bacteria 3
design: Total RNA was extracted from tissues using the CTAB method and genomic DNA was removed, and high-quality RNA samples were used for subsequent library construction. rRNA was removed using the RiboCop rRNA Depletion Kit for Mixed Bacterial Samples (lexogen, USA), mRNA was randomly broken into small fragments of about 200bp, mRNA was used as a template, Double-stranded cDNA was synthesized by reverse transcription using random primers. When the second strand of cDNA was synthesized, dUTP was used instead of dTTP. The synthesized double-stranded cDNA was complemented to the flat End by End Repair Mix, phosphorylated at the 5 'end, and added an A base at the 3' end to connect the Y-shaped sequencing linker. The second strand of cDNA containing dUTP was then eliminated with UNG enzyme, thus allowing the library to contain only the first strand of cDNA. RNA library construction was performed using Illumina Stranded mRNA Prep, Ligation from Illumina (San Diego, CA). Using illumina sequencer (NovaSeq6002) RNA - seq double-end sequencing.