Shotgun metagenomic DNA WGS of human disease state stool sample
Raw stools were thawed at room temperature and homogenized using a BeadBeater (BioSpec Products, Inc., Bartlesville, OK) for 3 min. DNA was extracted from a homogenized stool mix using the QIAamp DNA stool minikit (Qiagen, Inc., Limburg, Germany) and evaluated for purity and concentration using a NanoDrop 2000 device (Thermo Scientific, Inc., Waltham, MA) and Agilent QuBit device (Life Technologies, Inc., Carlsbad, CA). Extracted whole stool DNA was then sheared to 450 bp using a Covaris M220 sonicator and prepared for sequencing using the SPRIworks high-throughput sample preparation kit (Beckman Coulter, Inc., Brea, CA) and the NEBNext Ultra DNA library prep kit (New England BioLabs, Inc., Ipswich, MA).