16S RNA microbiota of mouse: germ-free stool
Microbial DNA was extracted using ZR Fecal DNA MiniPrep (Zymo Research) and purified with AMPure XP Beads (Beckman Coulter). The v3  v4 region of the 16S rRNA gene was first amplified by non-barcoded 341F and 805R primers and purified by AMPure XP Beads (Beckman Coulter). Followed by a second PCR amplification with Nextera XT index primers (Nextera XT index kit FC-131-1001 or FC-131-1002). The barcoded PCR amplicons were purified with AMPure XP Beads, quality checked on an Agilent 2100 BioAnalyzer (Agilent Technologies, Palo Alto, Calif., USA) prior to pooling.