title: Viral metagenome of Marthasterias glacialis
design: 1 g of preserved or frozen tissue was homogenized in 25 mL of 0.02um –filtered PBS in a NutriBullet blender. The homoegante was centrifuged briefly at 3,000 x g to remove large tissue debris and ossicles, and the supernatant filtered through 0.2um Durapore filters to remove cellular debris. The filtrate was precipitated with PEG-8000 (1 g/ml) overnight. The filtrate was then centrifuged at 22,000 x g for 30 min, supernatant decanted, and the pellet resuspended in 1 ml 0.02 um filtered PBS. The resuspension was filtered through a 0.2 µm Acrodisc PES filter. Samples were then treated with DNaseI, RNAse One and Benzonase for 3 h. Nuclease activity was stopped by the addition of x vol ETDA (25 mM). Viral DNA in the purified suspension was extracted using the Zymo Viral DNA kit. The DNA was amplified prior to sequencing using the WGA2 kit V2 (GE Biosciences) before submission to the Cornell Biotechnology Resource Center (BRC) for library preparation and sequencing