16S rRNA sequencing of fecal samples from adolescent Homo sapiens
Stool sample DNA was isolated with the Zymo Research Quick-DNA Fecal/Soil Microbe Kit (catalogue # D6010). After DNA standardization, the V4 region of the 16S rRNA gene was amplified with the New England Biolabs LongAmp Taq PCR kit (catalogue # E5200S) using primers and barcodes with slight modifications from the original Caporaso primers. Standardized amplicon libraries were pooled and submitted for 2  250 bp paired-end sequencing on the Illumina MiSeq System at the University of Alabama at Birmingham Genomics Institutional Research Core. All samples were sequenced on the same Illumina run to reduce batch effects.