Different approaches have been utilized to determine viral production in aquatic sediments, and a standardized protocol is not available yet. The protocol described here illustrates the procedures that have been optimized for determining viral production deep-sea sediment samples (Dell’Anno et al. 2009). This protocol is based on the dilution technique (i.e., the dilution of the sediment sample with virus-free seawater; Mei and Danovaro 2004) and allows quantifying directly any increase in viral abundance over time without using conversion factors.