GSM7908432: OVX.VEH4; mouse gut metagenome; OTHER
Colon flush was performed of the excised whole colon using sterile PBS. Bacterial genomic DNA was isolated from colon contents using QIAamp DNA Stool Mini Kit (Qiagen, Valencia, CA) and quantified using a Qubit 4 Fluorometer (ThermoFisher, Waltham, MA). Purified DNA was used in the library preparation for 16S ribosomal RNA sequencing using 16S Metagenomic Sequencing Library Preparation Kit (Illumina, San Diego, CA) according to the manufacturer's instructions. Briefly, the purified DNA was amplified by PCR using bacterial/archaeal sense 319 F primer and anti-sense 806 R primer to amplify the V3-V4 hypervariable regions of 16S rRNA. After clean-up, the amplified PCR products were then amplified again by PCR using dual indices and sequencing adaptors for 16S rRNA sequencing. Purified DNA was used in the library preparation for 16S ribosomal RNA sequencing using 16S Metagenomic Sequencing Library Preparation Kit (Illumina, San Diego, CA) according to the manufacturer's instructions.