WGS from human stool: Male
Fecal samples were flash frozen in dry ice, stored in -80 degree Celsius ultrafreezer, and transported to sequencing center in dry ice. DNA extraction using the MoBio PowerFecal DNA Isolation kit25 was performed at the sequencing center. Double-stranded DNA (dsDNA) concentration and quality was assessed using NanoDrop and the Qubit dsDNA Broad Range (BR) DNA Assay Kit26, respectively. DNA was diluted for library preparation using the Illumina Nextera XT Library Prep Kit, and 1 ng from each sample was fragmented and amplified using Illumina Nextera XT Index Kit primers. Amplified DNA was then cleaned using Agencourt AMPure XP beads, resuspended in buffer, and tested again for concentration, quality, and fragment size distribution on a bio analyzer using the Agilent High Sensitivity DNA Kit. DNA libraries were brought to the same nM concentration, pooled, and denatured with 0.2 N NaOH prior to loading on an Illumina MiSeq Reagent Kit v3 and sequencing on the Illumina MiSeq platform.