FIELD: chemistry.
SUBSTANCE: claimed invention relates to the field of biotechnology and deals with a method of amplification and detection of nucleotide sequences in a reaction mixture and a set, used in the said method. The claimed method includes providing an analysed sample, reagents for realisation of amplification by a polymerase chain reaction (PCR) and, at least, four double-labelled probes. Each of the probes is specific to its nucleotide sequence to be detected, bears a fluorescent label and a respective fluorescence quencher, with both probes bearing similar labels, and probes, bearing similar labels, differ by a temperature of melting (Tm). After that, amplification of the nucleotide sequences and an analysis of a melting curve are performed by means of the double-labelled probes in the reaction mixture.
EFFECT: solution can be applied in the multiplex PCR analysis in real time in diagnostics of bioreagents.
9 cl, 9 dwg, 21 tbl, 2 ex
