Bacterial dna extraction kit

Dry B. striata tubers were bought from Hubei Zexi Traditional Chinese Medicine Technology Co., Ltd. (Qichun, Hubei, China). The authenticity of medicinal materials was identified by Dr. Xiongjie Sun of Hubei University of Chinese Medicine. Vitamins, yeast extract, peptone, bacterial DNA extraction kit, bile salts, and L-cysteine were obtained from Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China). Pepsin, α-amylase, pancreatin, trypsin, and SCFAs standards (Acetic acid, propionic acid, butyric acid, pentanoic acid, and indole) were bought from Shanghai Aladdin Biochemical Technology Co., Ltd (Shanghai, China). All materials were of standard analytical grades.

Single colonies were inoculated into an anaerobic liquid medium and incubated anaerobically overnight. Following the operation manual procedures, genomic DNA was extracted from the bacterial solution using the Bacterial DNA Extraction Kit (Solarbio Technology Co., Ltd., Beijing, China) and stored at−20 °C. The extracted DNA was used as a template for five-fold polymerase chain reaction (PCR) amplification of the toxin genes tcdA, tcdB, and CDT, and the 16S rRNA following the method recommended by Cheng et al. (31 (link)). PCR amplification primer sequences were seen in Table 1. Additionally, A 1.8 kb at the 3' terminal of the tcdA gene was examined for deletion. The tcdC gene was also amplified and detected using the method proposed by Curry et al. (34 (link)). The sequence results were compared to the tcdC gene standard sequence (NC_009089.1, a gene bank accession number for tcdC gene sequence of Clostridioides difficile 630) to identify any gene deletion.