title: Virome of feces from CHD patients
design: For viral metagenomic analysis, 43 fecal specimens were collected from patients with coronary heart disease at Minhang Central Hospital of Shanghai. Study participants were 50 to 85 years old.  Each fecal specimen was resuspended with ten times the volume of DPBS and vibrated vigorously with an oscillator for 5 minutes. The supernatants were then collected after centrifugation (10 min, 15,000  g). 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. The remaining total nucleic acid was then isolated using QIAamp MinElute Virus Spin Kit according to the manufacturer's protocol. The 43 fecal samples from patients with coronary heart disease were randomly pooled into eight specimen pools five of which contained nucleic acids from five specimens, while three included nucleic acids from six specimens. These eight libraries were constructed using Nextera XT DNA Sample Preparation Kit (Illumina) and sequenced using the MiSeq Illumina platform with 250 bases paired ends with dual barcoding for each pool.