Search academic texts

Information × Registration Number 0214U006968, 0113U006053 , R & D reports Title The elaboration of the kits for DNA analysis of genomic rearrangements and epigenetic alterations, causing most common monogenic hereditary diseases of central and peripheral nervous system popup.stage_title Head Livshіts Ludmila Avramovna, Registration Date 17-04-2014 Organization Institute of Molecular Biology and Genetics of NASU popup.description2 The aim is to create prototypes (laboratory samples) of test kits for DNA analysis of genomic rearrangements (gene mutations) and epigenetic defects (methylation) in human genome that cause the most common hereditary determine diseases with severe disability associated with neurodegeneration of the central nervous system (Fragile X syndrome, Huntington's chorea, Prader-Willi and Angelman syndromes). In the second stage we optimized the components of the reaction mixtures, the temperature-time PCR profiles and the electrophoretic conditions for (a) hypermethylation status of fragile X sites and CGG-repeat expansion in FMR1 and FMR2 gene analyzes in males; (b) accurate number of FMR1 CGG-repeat determinination in females; (c) differential determination of hypermethylation status or deletions in 15q11-q13, (d) number of CGG and CCG-repeats determinination in IT15/HD. The laboratory samples of diagnostic kits (prototype test systems) were tested in DNA samples of male patients with a clinical diagnosis of fragile X syndrome (n = 20) and non-syndromic X-linked intellectual disability (n=40), Huntington's chorea (n = 20), Prader-Willi and Angelman syndromes (n = 35) as well as in DNA samples of patients' relatives and healthy control group. We selected the DNA samples of individuals with genetic alterations and normal genotypes, that were determined by Sanger sequencing. These samples are used as a control for the rest of the sample genotyping and verification of test results. The collection of representative DNA samples with accurately determined number of repeats in FMR1 and IT15 genes, with trinucleotide expansions in the FMR1 and FMR2, with hyper-and hypomethylation in FRAXA, FRAXE, FRAXF chromosomal sites, with impaired methylation and/or deletions of maternal/paternal alleles as well as control DNA samples with normal patterns of methylation 15q11-q13 chromosome region. DNA collection includes 25 control (reference) DNA samples. Product Description popup.authors Бичкова Г.М. Грищенко Н.В. Гулковський Р.В. Демидова Г.С. Корпан Я.І. Котенко О.В. Кравченко С.А. Крикливий І.А. Кучеренко А.М. Лівшиць Г.Б. Нечипоренко М.В. Пампуха В.М. Потужня О.О. Чабанова М.В. Чернушин С.Ю. popup.nrat_date 2020-04-02 Close