16S metagenomic sequencing of mus musculus fecal microbiota:  Subect 2025, Day 70, Base Diet Lean, Dietary Supplement Cellulose, Post-Surgery, Supplementation Ongoing
Fecal pellets were freshly harvested from mice after scruffing and immediately frozen at -80¼C.  DNA was extracted using the ZR Fecal DNA Extraction Kit (Zymo Research) as directed by the manufacturer.  16S ribosomal DNA (rDNA) was amplified with Phusion High-Fidelity polymerase (Thermo Scientific, Waltham, MA) and dual indexed primers specific to the V3-V4 hypervariable regions (319F: 5Õ ACTCCTACGGGAGGCAGCAG 3Õ; 806R: 3Õ ACTCCTACGGGAGGCAGCAG 5Õ)68.  Amplicons were pooled and paired-end sequenced on an Illumina MiSeq.