title: 16s rRNA of Homo sapiens : human nasal microbiome
design: Fecal samples were collected using a Commode Specimen Collection System, oral samples were collected using saliva collection tubes (Access-genetics, Eden Prairie, MN) and nasal samples were collected using ESwab (Copan Diagnostic, Murrieta, CA). All samples were then stored at -80C prior to processing. DNA was extracted from the fecal, oral and nasal samples using DNeasy PowerLyzer PowerSoil Kit (Qiagen, Germantown, MD) 16S rRNA regions (V3-4) were amplified as previously described23. DNA library for sequencing was prepared by Illumina MiSeq; we used the Nextera XT Index kit to attach dual index adapters. Each library was prepared by diluting the samples to 5 ng/Lm and equal volumes were mixed to 4 nM. We quantified the DNA concentration by Qubit (Thermo Fisher Scientific, USA). We carried out the library preparations according to the 16S library preparation protocol of Illumina (Illumina, San Diego, Cam USA), and sequenced the libraries using the MiSeq Reagent Kit v3 (600 cycles) for 300-bp pair-ends.