Intestinal microbial composition
MP
AMPLIFICATION
Fecal samples were collected before and after the experiment and were used to analyze changes in the composition of the gut microbiota. The resolved bacteria were extracted using the feces genome extraction kit from USA MP Company. After extraction, PCR was performed for the V3~V4 region of bacterial 16S rDNA. The PCR amplification system was as follows: 25 µL 2 × Premix Taq, 1 µL upstream primer 341F, 1 µL downstream primer 806R, 1 µL genomic DNA template, and 22 µL ddH2O. Amplification conditions were as follows: 95°C for 8 min; 95°C for 35 s, 52°C for 35 s, and 72°C for 40 s, 30 cycles; 72°C for 8 min. The gel was recovered using the ultra-thin agarose-gel purification and recovery kit from Beijing Tian Gen Biochemical Technology Co., LTD., and the purified fecal DNA was mixed according to the equal quality library samples and then sequenced by machine. QIIME1 software was used for data analysis after the microflora was disembarked.