METHOD OF DETECTING MUTATION P369ins IN GENE CYP1B1 Russian patent published in 2010 - IPC C12Q1/68 

FIELD: medicine.

SUBSTANCE: claimed is method of detecting new mutation in associated with primary congenital glaucoma (PCG) gene CYP1B1, which represents insertion of trinucleotide CTC, accompanied by appearance of additional proline residue in position 369 (P369ins) of human cytochrome P450 protein. Method includes PCR-amplification of fragment (349 nucleotide pairs) of DNA of 3-rd exon of gene CYP1B1 by means of direct and reverse primers 5'-CACCAAACAGGTATCCTGATGTG-3' and 5'-ATCACTCTGCTGGTCAGGTC-3'; carrying out heteroduplex analysis of PCR product, which includes its mixing with formamide in ratio 1:3, denaturation of obtained sample in boiling water, cooling to room temperature, DNA electrophoresis in 8% vertical polyacrylamide gel with further visualisation by staining with silver nitrate. Conclusion about presence of mutation in heterozygote state is made by formation of additional bands in gel, which have electrophoretic mobility of DNA fragments with length 420 and 450 pn, which are not detected in DNA samples, which do not contain P369ins.

EFFECT: method improvement.

1 dwg, 2 ex