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Information × Registration Number 0221U104432, 0116U005014 , R & D reports Title To research the regularities of development of oocytecumulus complexes of pig and receiving embryons from them in the culture in vitro depend on different amplitude and period of osci llation of temperature and concentration of pH popup.stage_title Head Denysiuk Pavlo V, Registration Date 23-03-2021 Organization Institute of Pig Breeding and Agro-Industrial Production of the National Academy of Agrarian Sciences of Ukraine popup.description2  A method of quantitative separation of oocyte-cumulus complexes into components has been developed, which provides for their preliminary fixation in a device of its own design. The patent protected by a utility model has shown high efficiency during experimental verification. Experimental verification of the design of the oligonucleotide primers MITPROF and R15615C showed the possibility of obtaining a specific product of amplification of porcine mitochondrial DNA with a size of 105 nucleotide pairs, which corresponds to the optimal size for quantitative real-time PCR. It was found that the use of oligonucleotide primers MITPROF and R15615C allows to obtain a specific product of amplification of mitochondrial DNA from a single pig oocyte. Real-time quantitative PCR to determine the relative concentration showed that the concentration of cumulus and oocyte DNA was 0.000083 and 0.000757 μg / μl, respectively, corresponding to the number of mitochondria in the oocyte of approximately 114,914 mitochondria. Real-time quantitative PCR to determine the relative concentration showed that the DNA concentrations in cultured oocytes 44, 46 and 48 h ranged from 0.000081 to 0.000083 μg / μl, did not differ significantly. Thus, it was found that in oocytes isolated from oocyte-cumulus complexes of the largest follicles, in which oocytes reach full size, the mtDNA content remains unchanged during meiotic maturation in vitro. According to the results, an article was published, which was included in the scientometric database of Web of Sciences and four patents for a utility model were obtained. Product Description popup.authors popup.nrat_date 2021-03-23 Close