Метод анализа числа копий GC-богатых повторяющихся последовательностей генома в составе поврежденной ДНК. Определение увеличенного содержания рибосомных генов в циркулирующей внеклеточной ДНК лиц с длительным стажем курения табака
2016
Detection of copy number variations of GC-rich repeated sequences in the structure of damaged DNA by PCR method results in significant understatement of their quantity in genome. We have described the method for analysis of the copy number variations of GC-rich repeat of human genome (mitochondrial, ribosomal and telomeric repeat) in damaged DNA. Comparing to others, these sequences of genome are mostly damaged in extracellular DNA, in DNA of the old cells, in long-stored DNA-samples and in oxidized DNA. The technology is based on the method of non-radioactive quantified dot hybridization with biotin-labeled DNA-probes and on the software designed specially to calculate the copy number of ribosomal genes (genes for rRNA) in extracellular DNA circulating in blood plasma of smokers’ and non-smokers’ humans. We have shown that fragments of GC-rich ribosomal repeat accumulate in extracellular DNA of chronically smoking people, which can be a sign of the intensification of necrotic processes in cells and of the chronic pathology development.
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