METHOD OF DETERMINING HUMAN GENOTYPE ON POLYMORPHISM IN TYPE II COLLAGEN GENE COL2A1 CA (RS1635529)

摘要

FIELD: biotechnology.;SUBSTANCE: invention is a method of determining the human genotype on polymorphism in type II collagen gene COL2A1 CA (rs1635529). This method can be used in diagnostics of periodontal disease in the oral cavity. The method comprises taking of the melting curves with fluorescently labelled allele-specific oligonucleotide samples. The pair of primers common to all alleles is used, fluorescently labelled allele-specific oligonucleotide samples different for each allele and a universal oligonucleotide labelled with fluorescence extinguisher of the following nucleotide composition: COL2_29s TCTTGCCCAAAGGCTAGGCGGA, COL2_29a CCAGAGGGCGAGAAAGAAACGA, COL2_29p1 AAAAAGGAAAGTTTATC-FAM, COL2_29p2 AAAAAGGAACGTTTATC-HEX, COL2_29pq BHQ 1-TTTGGATTTTCACTCTCTT-3'-(P), where FAM is a fluorescent dye FAM, HEX is a fluorescent dye HEX, BHQ1 is dark fluorescence extinguisher attached to the 5'-terminal nucleotide. The sample refers to homozygote or heterozygote for this allele by analysis of DNA melting curves forms, namely, the maximum of the first derivative of the fluorescence graphs.;EFFECT: proposed invention enables to carry out studies more accessibly to determine the polymorphism in the type II collagen gene COL2A1 of human genotype.;1 dwg, 2 tbl