title: shotgun metagenomic sequencing of human gut metagenome
design: DNA was extracted from ~200 mg of feces by the QIAamp Fast DNA Stool Mini Kit (Qiagen) in combination with bead beating on the FastPrep-24 System (MP Biomedicals). The shotgun genomic DNA libraries were constructed from 200 ng of DNA after sonication with a Covaris ME220 (Covaris) to an average fragment size of 300bp with the Hyper Library Preparation Kit from Kapa Biosystems. Libraries were pooled in equimolar concentration and size selected on a 2% agarose gel for fragments 220bp to 280bp in length. The pool was further quantitated by qPCR. The pooled barcoded shotgun libraries were loaded on a NovaSeq lane for cluster formation and sequencing at a length of 150nt from each side of the DNA fragments.