METHOD FOR DETERMINING EXPRESSION LEVEL OF GENE CODING 1L-1β IN RABBIT EYE TISSUES (ORYCTOLAGUS CUNICULUS), AND KIT FOR IMPLEMENTATION THEREOF

摘要

FIELD: biotechnology.;SUBSTANCE: group of inventions relates to biotechnology, specifically immunology and molecular biology, and a method of determining the level of transcription of a gene coding IL-1 β in rabbit eye tissues (Oryctolagus cuniculus) using RT-PCR in real time. Method comprises selecting original primer pairs for IL-1 β and GAPDH, providing specific amplification without formation of side structures, determining reaction mixture components - concentrations of a pair of forward and reverse primers, setting optimum elongation temperatures, denaturation and annealing (using a gradient method) and the number of amplification cycles; use of one normal gene GAPDH, as most universal, stably expressed practically with all body cells. A kit containing original oligonucleotide primers for the IL-1 gene β rabbit Oryctolagus cuniculus, original oligonucleotide primers for the GAPDH gene, PCR real-time PCR reaction for amplification of IL-1 gene β, PCR reaction mixture in real time for amplification of GAPDH gene.;EFFECT: invention can be used in experimental ophthalmology, as well as at preclinical studies, particularly when testing anti-inflammatory drugs.;2 cl, 3 dwg, 1 tbl