16S rRNA feces sequencing of prediabetic subjects with obesity after antibiotic therapy - methane non-eradicated subject
Sequencing of the V4 region of the bacterial 16S rRNA gene was performed using the barcoded primer set 515f/806r and following protocols from the Earth Microbiome Project. 240 ng of DNA per sample were pooled and purified using QIAquick PCR purification kits. The PCR pool was quantitated using the KAPA Library Quantification Kit and diluted to a concentration of 4 nM, denatured, and then diluted to a final concentration of 4 pM with a 15% PhiX spike