An anaerobic cultivation to study the effect of TMA on human gut microbiota

We collected 11 fresh faecal samples from 11 healthy individuals who had taken no antibiotics for three months prior to this study. Then, the faecal dilutions (10-5 dilution) were inoculated into Brain Heart Infusion Broth (BHI broth) and BHI broth + 4 g/L TMA (v/v 1:25, each 10-5 dilution was inoculated in triplicate) inside an anaerobic chamber. After anaerobic incubation for 7 days, DNA was extracted and amplified with barcoded V4 primers. The PCR products were sequenced using the Illumina Hiseq PE250 sequencing strategy.