Various options and features of equipment for real-time PCR were considered, recommendations for the selection of an amplifier were given. The features of systems for fluorescent registration of DNA accumulation are analyzed. Key factors determining oligonucleotide sequence selection and amplification program parameters are considered. Attention is paid to the preparation of samples and the features of the analysis of the data obtained, which is necessary to obtain the most reliable results. Separate chapters are devoted to the use of real-time PCR to solve various problems: determining the level of transcript representation, viral load, nucleotide polymorphism, and the relative content of nucleic acids using the example of GMOs.