title: 16S rRNA seq of microbiota: feces
design: For streptomycin-treatment experiments, fecal samples were stored at -80 C and DNA extraction was performed as previously described16. In brief, DNA was extracted using a combination of the QIAamp Fast DNA Stool Mini Kit (Qiagen) following the manufacturers instructions, mechanically disrupted with a tissue blender, and eluted to a final volume of 100 L in ATE buffer.  For each sample, the 16S rRNA gene was amplified using the Earth Microbiome Project-recommended 515F/806R primer pairs (V3-V4 regions) under the following PCR cycling conditions: 94 C for 3 min, 35 cycles of 94 C for 60 s, 50 C for 60 s, and 72 C for 105 s, with an extension step at 72 C for 10 min57,58.  After library prep, 2x250-bp sequencing was performed at the IGC Genomics Unit using an Illumina MiSeq Benchtop Sequencer.
identifiers.namespace: SUB6567942
identifiers.id: sample214