GSM7233245: FM2-5; Clostridioides difficile; OTHER
Individual clones of bacteria, recovered from faecal or tissue associated material that showed different morphology on ChromID plates, where subject to at least two rounds of clonal selection. Genomic DNA was isolated from a 20 ml culture grown anaerobically for 18 h in tryptic soya broth (TSB). Bacterial cells were initially disrupted enzymatically by resuspending the pellet in lysis buffer (20 mM Tris-Cl, pH8.0, 2 mM Na EDTA, 1.2 % Triton X-100, lysozyme 200 mg ml-1), and incubated at 56 oC for 90 min. The DNA was recovered using the DNeasy Blood and Tissue Kit (Qiagen), following manufacturer's instructions. Genomic DNA recovered from R20291, FM2.5, FM2.5varA, FM2.5varB were sequenced by MicrobesNG (Birmingham UK) using Illumina next-seq 500 with 250bp paired read length.