title: 16s rDNA amplicon of mouse feces (control +  allicin group)
design: All fecal samples were aliquoted  and stored at -80 degrees Celcius for further examination. Fecal genomic DNA  was extracted using the QIAamp PowerFecal DNA Kit according to the  manufacturer's instructions. Two-step polymerase chain reaction (PCR) workflow  was performed for library preparation under procedures described in the  Illumina 16S sample preparation guide. The 16S rRNA gene V3-V4 region was  amplified and sequenced on the Illumina Miseq platform for paired-end  sequencing (2 x 300 bps). The 16S rRNA gene V3-V4 region was amplified using  a primer overhanging adapter (forward =  5-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGCCTACGGGNGGCWGC    AG-3 and reverse = 5 GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGACTACHVGGGT    ATCTAATCC-3). Dual indices and Illumina sequencing adapters were attached  through PCR by using a Nextera XT Index Kit according to the manufacturers  instructions. After each PCR process, PCR cleanup was performed using AMPure  XP beads to purify V3-V4 amplicon from the free primer and primer dimer. The  sizes of PCR products were verified using the Bioanalyzer DNA 1000 chip.  Library quantification was performed for quality control before sequencing by  using the Agilent Technologies 2100 Bioanalyzer. The pooled libraries were  then sequenced on the Illumina Miseq platform with v3 reagents for paired-end  sequencing (2  300 bps).