Nucleic acid metabolism processes such as the synthesis and degradation of DNA and RNA are the basic metabolic units to maintain the growth and development, metabolism, genetic variation and aging, and widely participate in the whole process of the body’s life activity. The enzyme activity related to nucleic acid metabolism is crucial for maintaining the stability of the intracellular environment, and the change of the activity may cause the occurrence and development of many diseases. The enzymes related to nucleic acid metabolism have become important targets for the study of various diseases and are indispensable tools in the field of biotechnology and bioengineering, such as polymerase chain reaction, site-directed mutagenesis, molecular cloning and DNA sequencing. Therefore, nucleic acid metabolism is the basis of all nucleic acid studies and related life science researches. In this paper, we introduce the common methods of enzymatic analysis of nucleic acid metabolism, and focus on the simple and fast real-time fluorescence method, classify and compare them according to their principles, development history and applications, and also prospect the future study and development of tools for enzymatic analysis of nucleic acid.