METHOD FOR PROTECTION AGAINST CONTAMINATION OF REACTION MIXTURES FOR PCR Russian patent published in 2023 - IPC C12Q1/6848 

FIELD: biotechnology.

SUBSTANCE: method for protection against contamination of reaction mixtures for PCR is described. PCR is carried out with the addition of 2'-deoxyuridine-5'-triphosphate, uracil DNA glycosylase and DNA glycosylase Fpg, Nei. Results are detected using an intercalating dye, TaqMan probe, or gel electrophoresis, oligonucleotide primers, and a DNA or RNA template. The absence of contamination with products of previous PCR runs is determined by the absence of amplification in PCR samples without a template. Decreased concentration of DNA template from 2'-deoxythymidine-5'-monophosphate is determined by an increase in Cq values or by a decrease in the intensity of PCR product bands on electropherograms in samples containing UG and DNA glycosylases Fpg, Nei, compared with samples with UG without Fpg, Nei.

EFFECT: invention makes it possible to increase the effectiveness of PCR protection against contamination by introducing single-strand breaks in DNA fragments containing apyrimidine sites. When Fpg and Nei are added to the reaction mixture for PCR, the concentration efficiency of PCR products with dUMP decreases by 500–1,000 times compared with control samples with UG without additional enzymes.

2 cl, 3 dwg, 1 tbl, 5 ex