Y21_fecal
For each fecal sample, 0.25 g aliquot was extracted using Qiagen Fast Stool DNA isolation kit with slight modification. In the first lysis step with InhibitEx Buffer, 0.4g of silica beads (0.1mm 0.2g and 1.0 mm 0.2g) was added into the sample prior to homogenization in FastPrep-24 (MP Biomedicals) for 1 minute at 4 m per sec. Library preparation was performed according to Illuminas TruSeq Nano DNA Sample Preparation protocol. The samples were sheared on a Covaris E220 to ~450bp, following the manufacturers recommendation, and uniquely tagged with one of Illuminas TruSeq HT DNA barcodes to enable sample pooling for sequencing. Finished libraries were quantitated using Promega's QuantiFluor dsDNA assay and the average library size was determined on an Agilent Tapestation 4200. Library concentrations were then normalized to 4nM and validated by qPCR on a QuantStudio-3 real-time PCR system (Applied Biosystems), using the Kapa library quantification kit for Illumina platforms (Kapa Biosystems). Libraries were then pooled at equimolar concentrations and sequenced on an Illumina HiSeq2500 sequencer in rapid mode and a read-length of 251bp paired-end (Illumina V2 Rapid sequencing reagents).