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4.9. Microbiome Analysis by 16S rRNA Gene Sequencing

CW Chenyang Wu
TZ Tingting Zheng
HC Huan Chen
PZ Peizhi Zou
MZ Mengxue Zhang
JW Jinrui Wang
NL Nan Li
YZ Yun Zhang
YL Ying Li
ZD Zhengqi Dong
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Feces of all mice were collected for further analysis and stored at −80 °C, Microbial community genomic DNA was extracted by utilizing the E.Z.N.A® soil DNA Kit (Omega Bio-Tek, Norcross, GA, USA). The hypervariable region V3–V4 of the bacterial 16S rRNA genes was amplified by an ABI GeneAmp® 9700 PCR thermocycler (ABI, CA, USA). Purified amplicons were pooled in equimolar and paired-end sequenced on an Illumina MiSeqPE300 platform/NovaSeqPE250 platform (Illumina, San Diego, CA, USA) according to the standard protocols by Beijing Expand biotech Science & Technology Co.Ltd. (Beijing, China). The taxonomy of each OTU representative sequence was analyzed by RDP Classifier (version 2.2) against the 16S rRNA database (egg. Silva v138) using a confidence threshold of 0.7.

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Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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