Raw data for supplementary fig S1-submission 619878

Published: 23-11-2020| Version 2 | DOI: 10.17632/4gfs7mmm2c.2
Chien-Hsiung Pan


Raw data for supplementary fig S1-submission 619878 (Frontiers in Microbiology; Nov 20, 2020)

Steps to reproduce

1. Gut microbiota analysis Stool from individual mice were weighed and homogenized in 0.5 ml of PBS. Fecal DNA was purified by the MPbio DNA/RNA isolation kit based on the direction of manufacture. The bacteria 16S rRNA gene was determined by illumine sequencing. In brief, the bacterial 16S rRNA genes were amplified by KAPA High-Fidelity PCR Master Mix (KAPA BIOSYSTEMS) using the specific primers targeting the V3 and V4 hypervariable regions (341F-CCTACGGGNGGCWGCAG and 805R- GACTACHVGGGTATCTAATCC) with the barcodes. The PCR product was purified with QIAquick gel extraction kit (QIAGEN). Sequencing libraries were generated using Truseq nano DNA Library Prep Kit (Illumina, USA) following manufacturer’s recommendations and index codes were added. The library was sequenced on an Illumina Miseq platform to generate 300 bp paired-end reads. The operational taxonomic units (OTUs) generated from quality-filtered and non-chimeric reads were analyzed with SILVA 132 OUT collection for bacterial OUT taxonomy assignment.