16S rRNA analysis of fecal microbiota

HL Haonan Li
YX Yujiao Xiang
ZZ Zemeng Zhu
WW Wei Wang
ZJ Zhijun Jiang
MZ Mingyue Zhao
SC Shuyue Cheng
FP Fang Pan
DL Dexiang Liu
RH Roger C. M. Ho
CH Cyrus S. H. Ho
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The experiments included extracting the total DNA from samples (n = 5 per groups) of the faeces. The data were analyzed on the free online Majorbio I-Sanger Cloud Platform. Total DNA was extracted according to the instructions of the E.Z.N.A.® SOIL Kit (Omega Bio-Tek, Norcross, GA, U.S.). The concentration and purity of DNA were measured using a NanoDrop 2000 spectrophotometer, and the quality of the DNA extraction was confirmed by 1% agarose gel electrophoresis. PCR amplification of the V3–V4 variable region was performed using 338F (5′-ACTCCTACGGGAGCAGCAG-3′) and 806R (5′-GGACTACHVGGGTWTCTAAT-3′) primers. The microbial composition was analyzed via 16S rRNA sequencing by Shanghai Majorbio Bio-pharm Technology (Shanghai, China) according to standard instructions.

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