title: 16S rDNA of mice: adult female feces
design: The total genomic DNA was extracted from the fecal samples using the E.Z.N.A. Soil DNA kit (Omega Bio-tek, GA) according to the manufacturer's protocols. The concentration and quality of DNA were checked using NanoDrop 2000 UV-vis spectrophotometer (Thermo Scientific, Wilmington, DE, USA). The V3V4 regions of the bacterias 16S rRNA were amplified with universal primers 338F (5-ACTCCTACGGGAGGCAGCAG-3) and 806R (5-GGACTACHVGGGTWTCTAAT-3) using a thermocycler PCR system (GeneAmp 9700 ABI, Carlsbad, CA, USA). The amplification procedures were conducted at 95 C for 3 min in the first place, followed by 27 cycles (denaturation at 95 C for 30 s, annealing at 55 C for 30 s, and elongation at 72C for 45 s) and a final extension (72 C, 11 min).