Detailed and comprehensive DNA interpretation and comparison protocols are needed to ensure reliable and consistent interpretation and comparison of DNA data from single source and mixed DNA samples. This document describes specific requirements for a laboratory’s DNA interpretation and comparison protocol(s) and provides direction for its development. These requirements include distinguishing single source data from mixed data, defining assumptions that may be used, defining limitations of the interpretation methods and determining when data are unsuitable for interpretation or comparison based on the laboratory’s internal validation studies, published scientific literature and other appropriate scientific resources where available. A requirement for a documented policy to ensure that evidentiary data are interpreted prior to comparison to known reference data is provided. The goal is for the laboratory to consistently produce reliable, repeatable and reproductible interpretations and conclusions that are supported by internal validation data and laboratory protocols.