16S rRNA gene sequencing of gut microbiota from time-restricted feeding and fecal microbiota transplantation mice

Time-restricted feeding (TRF) donor mice were allowed free access to food only between 10:00 and 18:00 each day for 2 weeks, whereas control (CTRL) donor mice had ad libitum access to food throughout the day. Fecal samples from donor mice were collected and used to prepare fecal microbiota suspensions. Recipient mice were pretreated with an antibiotic cocktail by oral gavage for 3 consecutive days, followed by daily oral gavage for 5 consecutive days with fecal microbiota suspensions derived from either CTRL or TRF donors or with the cell-free fecal supernatant prepared from CTRL donor fecal suspensions by centrifugation at 8,000 x g for 5 min. Fecal samples were collected from donor mice (CTRL and TRF) and recipient mice after treatment (FMT-CTRL, FMT-TRF, and FMT-Sup) for 16S rRNA gene sequencing to characterize gut microbial composition.