title: Metagenome of homo sapiens: infant gut
design: DNA was extracted from human fecal samples in two stages. First, approximately 50 mg of fecal material and 650 L MBL lysis buffer from the Qiagen PowerMicrobiome DNA/RNA EP Kit were added to Lysis Matrix E (LME) tubes (MP Biomedicals).  LME tubes were transferred to a Precelleys 24 Tissue Homogenizer (Bertin Technologies) and fecal samples were homogenized, centrifuged, with the resultant supernatant transferred to a deep-well 96-well plate. The second stage consisted of DNA isolation from the above supernatant using the Qiagen MagAttract PowerMicrobiome DNA/RNA EP Kit on an Eppendorf automated liquid handling system as detailed by the manufacturer. Total gene content of the microbiome was assessed through shotgun metagenomic sequencing. Metagenomic libraries were constructed from 100 ng of DNA as starting material using the Illumina DNA Prep kit. Illumina DNA/RNA UD Indexes were used to add sample-specific sequencing indices to both ends of the libraries. An Agilent 4200 TapeStation system with High Sensitivity D5000 ScreenTape (Agilent Technologies, Inc) was used to verify quality and assess final library size. Metagenomic libraries were normalized and pooled at an equimolar concentration. Final pools were sequenced on a NovaSeq X sequencer using a paired-end (150x150) NovaSeq 25B flow cell across ten lanes. (Illumina, Inc).