Abstract: A great variety of nucleic acid reference materials have been developed to improve the accuracy of nucleic acid measurement. A reference material is a “measuring instrument” that can ensure the accuracy and traceability of measurement, with the functions of reproducing, preserving, and disseminating measurement values. Reference materials can provide a frame of reference for quality control of qualitative and quantitative detection processes for nucleic acid. Accurate and reliable certification methods are an important basis for the development of reference materials. This article provides details on several methods of measuring nucleic acid reference materials, with a focus on the principles and application characteristics of the methods and a discussion on influencing factors that may affect the measurement.