METHOD FOR POINT MUTATION DIAGNOSING IN NATIVE DNA USING GRAPHENE OXIDE Russian patent published in 2017 - IPC C12N15/00 

FIELD: biotechnology.

SUBSTANCE: for point mutations (substitutions, insertions, deletions) diagnosis in the native DNA, allele-specific PCR carried out with two fluorescently labeled forward primers complementary to DNA sequences of different alleles near the mutation site, and reverse common primer, with subsequent addition of a graphene oxide suspension (as a selective nanostructured fluorescence quencher) to PCR products and measurement of resulting solution fluorescence intensities in two fluorescence channels corresponding to the forward primers fluorescent labels. For comparison and obtaining of diagnostic results, negative control samples not containing DNA on PCR step are applied. Comparison of fluorescence intensities in the final DNA analyte sample solution in two fluorescence channels with the same intensities for negative control, allows to determine the analyzed DNA sample genotype. At the same time, for 4582insT mutation diagnosis in exon 25 of the CUL7 gene causing 3M syndrome in Yakuts, 5'-FAM-CAGGGGTCCTCAAGATTTCG-3'; 5'-ROX-CAGGGGTCCTCAAGATTCG-3' oligonucleotides are used as forward primers; 5'-GATGAGGCAGTTCAGAAGATTCC-3' oligonucleotide is used as a reverse primer.

EFFECT: development of a reliable method for point mutations diagnosis in the native DNA using graphene oxide as a selective fluorescence quencher, promising for development of methods for DNA diagnostics of hereditary and genetically predisposed diseases, pharmacogenetic tests.

2 cl, 4 dwg, 3 tbl