16S of synthetic sputum community
DNA purification, sequencing, and qPCR were performed by MR DNA Lab (Shallowater, TX). Briefly: DNA was purified from sputum homogenate after mechanical lysis with the MoBio Power Soil kit (MoBio, Carlsbad, CA). The 16S V4 region of the resulting DNA was amplified with 515F and 806R primers incorporating the barcode in the forward primer and subjected to Illumina sequencing(1). The sequence data were generated in a total of 6 MiSeq runs. Total 16S abundance in each sample was determined by qPCR using standard 515F/806R primers(1).
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