the regulations of systemic EVs on 16S rDNA gut microbiota after seizures
This project investigates changes in gut microbiota composition in a mouse model of epilepsy treated with exosomes (EXO). Fecal samples were collected from three groups - negative control (N), epilepsy model (PBS), and treatment (EXO) - at four time points (4 days, 7 days, 14 days, and 1 month post-induction). Samples were pooled from 3 cages per group/time point (expanded to 8 at 14 days) and stored at -80 C. Genomic DNA was extracted from 300 uL fecal supernatant using the Nucleic Acid Extraction and Purification Kit (Xiamen TreatGut Biotechnology Co., Ltd.). The 16S rRNA V4 region was amplified using primers 515F and 806R with the Human Gut Flora Genetic Detection Kit, followed by PCR purification, library pooling at 40 ng/sample, circularization, and sequencing on the MGI DNBSEQ-G99 platform generating paired-end reads. Bioinformatic analysis included read merging with FLASH, quality filtering with Cutadapt, OTU clustering at 97% similarity using USEARCH with UCHIME chimera removal, taxonomy assignment against SILVA 132 97 16S via RDP Classifier, and diversity metrics. Differential taxa were identified with LEfSe and indicspecies, and functional predictions were made using PICRUSt2. Data will be released upon publication.