16S of Mus musculus under following experimental condition: P016.8nxU in Mouse
Fecal DNA was extracted by bead-beating and quantified with Qubit assays (Life Technologies). Each fecal DNA template was normalized to 2ng/l and a PCR was performed to add indexes and amplify the DNA. The samples were quantified with Qubit dsDNA HS assays and pooled together into equal amounts. The pooled sample was purified with AmpureXP beads (Beckman Coulter) with a ratio of 1:1 beads to PCR reaction. Correct amplicon size and the absence of primer dimers were verified by gel electrophoresis. The 16S rDNA amplicon sequencing of the V4 hypervariable was performed with an Illumina MiSeq (paired-end 250bp).