The paper presents the results of studies on the development of metrological support for DNA sequencing. The sequencing of a standard sample of the human mitochondrial DNA was carried out according to the Sanger method on domestic capillary electrophoresis genetic analyzers Nanophore 05. The results of the work were used to validate standard reference material for human mitochondrial DNA sequence and to confirm the DNA sequencer as a measurement instrument.
DNA sequencing is an essential task of modern molecular genetic research. Its fundamental parameter is the accuracy of decoding. In this work, we determined the maximum lengths of DNA fragments, obtained by the Sanger method and sequenced with an accuracy of 99%, in a polymer based on linear poly(N,N-dimethylacrylamide) PDMA-6 for capillaries 50 cm long to the optical window of the detector of a Nanophore 05 domestic genetic analyzer and an imported GA 3500xL genetic analyzer. The results of DNA sequencing obtained by various analysis algorithms were compared. The parameters of the analysis algorithms were selected, ensuring best results of DNA sequencing in using the PDMA-6 polymer.
The heterozygosity of CANP3, ACTN3 , and GHR genes in specialized collections was studied using state-of-the-art DNA technologies for DNA analysis. A new dinucleotide deletion (AC) at the beginning of exon 21 was identified in five individuals with a heterozygous CANP3 gene. Analysis of polymorphism (1747 C T) of the ACTN3 gene demonstrated a positive association of allele C with high muscular performance. Real-time PCR assay of SNP1630 (A C) in the GHR gene suggested a putative negative association of allele C of this SNP with high muscular performance.
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