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METHOD FOR DETERMINING GENOTOXICITY OF XENOBIOTICS BASED ON ANALYSIS OF DAMAGE TO MITOCHONDRIAL DNA OF EARTH BUMBLEBEE (BOMBUS TERRESTRIS)
METHOD FOR DETERMINING GENOTOXICITY OF XENOBIOTICS BASED ON ANALYSIS OF DAMAGE TO MITOCHONDRIAL DNA OF EARTH BUMBLEBEE (BOMBUS TERRESTRIS)
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机译:基于对土蜂线粒体DNA损伤分析的异种药遗传毒性测定方法
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摘要
FIELD: biotechnology.;SUBSTANCE: invention relates to biotechnology. Method for determining the genotoxicity of xenobiotics based on an analysis of damage to mitochondrial DNA of earth bumblebee (Bombus terrestris). Method comprises feeding ground bumblebees (Bombus terrestris) with the tested xenobiotics throughout the day, sampling of biological material and isolation of DNA from pectoral muscles of the bumblebee, determination of the quantitative level of damage to mitochondrial DNA by real-time PCR at an elongation temperature of 66.1±0.5 °C for 5 minutes using Encyclo-polymerase corresponding to Encyclo-buffer and other standard reaction components with primer pairs having specific sequences, comparison of the quantitative level of damage to the mitochondrial DNA of the experimental and control groups. Higher level of damage in the experimental group is an indicator of the genotoxicity of xenobiotics. Sequences of primers for determining the amount of damage in mitochondrial DNA B. terrestris and the number of its copies:;
;Present invention can be used in testing various xenobiotics, including pesticides, for genotoxicity.;EFFECT: technical result consists in development of highly sensitive, highly reproducible and relatively inexpensive way to determine the genotoxicity of xenobiotics.;1 cl, 2 dwg, 1 ex
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