16S rRNA gene V4 amplicon sequencing of nasal lavage fluid from healthy child
Total DNA was extracted from nasal lavage fluid using the Bacteremia DNA Isolation Kit (MO BIO Laboratories Inc). The full length 16S rRNA gene was amplified for 15 cycles using primers 27F and 1492R. Subsequently, the 16S rRNA gene V4 region was amplified in triplicate for 15 cycles using primers 515F and 806R that included the P5 and P7 Illumina Adapters, respectively, and Index sequences. These reactions were pooled and sequenced using the 2300 bp paired-end Illumina MiSeq platform at the University of Wisconsin-Madison Biotechnology Center (Madison, WI).