The metagenomic studies on viral diversity of human fecal specimens. Homo sapiens

For viral metagenomic analysis, 100 fecal specimens were collected from newborns who were admitted to the newborn department of the Affiliated Hospital of Jiangsu University from Jan 2016 to Dec 2016. Each specimen was added 500 ul DPBS. And then each specimen was vortex for 5 minutes and repeated it three times. The supernatants were collected after centrifugation (10 min, 15,000 g). Then the 100 supernatants were randomly pooled into 10 specimen pools, each including 10 specimens. Supernatants were filtered through a 0.45-mm filter (Millipore) to remove eukaryotic- and bacterial cell-sized particles, and 200 ul of supernatant from each pool was then subjected to a mixture of nuclease enzymes to reduce the concentration of free (non-viral encapsidated) nucleic acids. Remaining total nucleic acid was then isolated using QIAamp MinElute Virus Spin Kit according to manufacturer's protocol. Ten libraries were then constructed using Nextera XT DNA Sample Preparation Kit (Illumina) and sequenced using the Illumina sequencing platform with 250 bases paired ends with dual barcoding for each pool.