SET OF OLIGONUCLEOTIDE PRIMERS AND METHOD FOR GENOTYPING SINGLE NUCLEOTIDE POLYMORPHISM rs8065080 IN HUMAN TRPV1 GENE

摘要

FIELD: biotechnology.;SUBSTANCE: technical solution relates to the field of biotechnology. The essence is a set of 3 primers for the determination of SNP rs8065080 in the TRPV1 gene, which have the activity of forward and reverse primers in the polymerase chain reaction and have the following structure;Forward1: 5’-TGTGCCGTTTCATGTTTGTCTTCG-3̓ (24 n.);Forward2: 5’-CTGTGCCGTTTCATGTTTGTCTTCA-3’ (25 n.);Reverse: 5’-TGAGGTAGGAGAATTGCTTGAACC-3’ (24 n.);Method for genotyping a single nucleotide polymorphism rs8065080 in the human TRPV1 1 gene, which consists in preparing reaction mixtures 1 and 2, containing components for carrying out the PCR reaction, to carry out AC-PCR, then the Forward1 primer and the Reverse primer according to claim 1 are added to the reaction mixture 1 , then the Forward2 primer and the Reverse primer according to claim 1 are added to the reaction mixture 2, then the analyzed DNA is added to the reaction mixture 1 and the reaction mixture 2, then the AC-PCR reaction is carried out for the reaction mixture 1 and the reaction mixture 2 according to the following program: 94°C – 3 minutes; further 33 cycles: 94°C – 30 sec, 64°C – 10 sec, 72°C – 20 sec; further 72°C – 2 min; then 4°С – ∞, at this the AC-PCR reaction is completed; then the amplification products are separated by electrophoresis in 1.5% agarose gel, then DNA is visualized by staining with ethidium bromide and subsequent UV irradiation. Identification of the rs8065080 single nucleotide polymorphism in the TRPV1 gene is a promising method for stratification of patients with a wide range of conditions accompanied by pain, such as various types of migraine, osteoarthritis, postherpetic neuralgia, diabetic, post-traumatic or postoperative neuropathic pain syndrome, which makes it possible to conduct an informed choice preventing the chronicity of pain in patients, taking into account the characteristics of the genome of a particular patient.;EFFECT: makes it possible to conduct an informed choice preventing the chronicity of pain in patients, taking into account the characteristics of the genome of a particular patient.;2 cl, 2 dwg, 1 tbl