RNA-seq of Clostridioides difficile ∆rpoN mutant against wild-type control
RNA-seq of Clostridioides difficile ∆rpoN mutant against wild-type control
The WT, ΔrpoN, and ::rpoN strains were inoculated into BHIS medium and cultured until the OD600 reached 0.6. The cultures were then centrifuged at at 13,500 × g for 10 minutes, the supernatant was removed, and the bacterial pellets of each strain were collected. The samples were transported with adequate dry ice in an foam box to Nanjing Personalomics Co., Ltd. for transcriptome sequencing. Total RNA was extracted from WT, ΔrpoN, and ::rpoN using a bacterial total RNA extraction kit (DP430, TIANGEN, Beijing). Once the samples were deemed satisfactory, the following steps were conducted for sequencing: (1) Removal of ribosomal RNA; (2) Enrichment and purification of mRNA; (3) Fragmentation of mRNA; (4) Construction of the sequencing library and quality assessment of the library.