title: Transcriptome, inflammatory and oxidative properties of Clostridium butyricum CB1002 clinical strain and its isogenic mutant CB1002 Δhbd
design: Transcriptome, inflammatory and oxidative properties of Clostridium butyricum CB1002 clinical strain and its isogenic mutant CB1002 Δhbd
library_construction_protocol: samples were subcultured on columbia cystein agar at 37°C for 48 hours, and then cultured in 10 mL BHI broth firstly and 20 mL BHI broth secondly overnight at 37°C prior to isolation. All cultures have been perfomed in an anaerobic chamber (Coy Laboratory Products Inc, Grass Lake, MI, USA) (v/v: 90% N2, 5% H2, 5% CO2). Clostridium butyricum cultures were incubated until it reached an optical density (OD600) of 1.1 for isolation at mid-exponential phase. Total bacterial RNA was then extracted and purified using ZYMO Direct-zol™ RNA Miniprep Plus R2070 extraction kit and treated with DNAse I according to manufacturer's instructions. A total of 100 ng RNA sample was used to construct each RNAseq library using Illumina stranded Total RNA Ligation Ribo0+ kit and subjected to ribosomal RNA (rRNA) depletion following manufacturer's instructions