Genetic Analyzer Nanofor 05 as a Measuring Instrument for DNA Sequencing

Volkov, A. A.; Volkov, I. A.; Plugov, Aleksandr G.; Kulyabina, E. V.; Melkova, Olga N.; Lavrov, Georgii S.; Bocharova, D. V.; Alekseev, Ya. I.

doi:10.1007/s11018-021-01897-2

Meas Tech

2021

DOI: 10.1007/s11018-021-01897-2

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Genetic Analyzer Nanofor 05 as a Measuring Instrument for DNA Sequencing

A. A. Volkov

I. A. Volkov

Aleksandr G. Plugov

et al.

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Cited by 2 publications

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“…When reading the section of the reference sample using a domestic genetic analyzer (Nanofor 05) based on capillary electrophoresis, a reading accuracy of 98.5% was achieved under conditions of repeatability and reproducibility based on the results from various laboratories [ 9 ].…”

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Metrological Support of Nucleic Acid Sequence Analysis

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Metrological support for the process of molecular genetic identification is proposed. An establishment of a nucleotide sequence is the main objective of the DNA/RNA analysis. Such nucleotide sequence provides key information about the role and function of nucleic acids in living systems, as well as the origin of biological objects during molecular genetic identification. The molecular genetic identification is based on the modern methods of studying DNA sequences, which are inherited by the living organisms from generation to generation. In some cases, comparison of DNA sequences is required to establish the authenticity of biological objects, as well as determine biological affiliation or kinship with other biological objects. The problems of metrological support of molecular genetic identification by Sanger DNA sequencing using fluorescent-labeled reaction terminators and capillary electrophoresis are considered. Methods and means of metrological support of the DNA/RNA analysis were developed, which include the determination of a nucleotide as a nucleic acid monomer, the use of a nucleotide sequence of nucleic acids used in certified methods and procedures, the use of a genetic analyzer as a measurement tool along with related auxiliary equipment, as well as the use of a standard sample of a fragment of human mitochondrial DNA has been developed and approved. A Nanofor 05 domestic genome analyzer was tested. The following two procedures were developed and certified: for measuring the nucleotide sequence of the section of the control region of mitochondrial DNA of fish (sturgeon and paddlefish families) by Sanger sequencing, and for measuring the nucleotide sequence of the 5' region of the COI gene of the mitochondrial DNA of aquatic biological resources by Sanger sequencing. Both measurement procedures are based on the use of fluorescent-labeled reaction terminators and capillary electrophoresis. The results are important for use in the DNA studies in the field of biological sciences, food industry, biotechnology, and DNA forensics.

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Metrological Support of Nucleic Acid Sequence Analysis

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Metrological support of sequence analysis of nucleic acids

Melkova

Kulyabina

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Metrological support for the process of molecular genetic identification is provided. An establishment of a nucleotide sequence is the general aim in nucleic acids analysis. A nucleotide sequence of DNA/RNA is the key information which gives an idea of the role and functioning of nucleic acids in living systems, as well as it allows establishing the origin of biological objects during molecular genetic identification. The molecular genetic identification is based on modern methods of research in the sequencing of DNA, which is inherited from generation to next generation of living organisms. DNA sequence comparison in some cases is mandatory to establish the authenticity of biological objects and to determine biological affiliation or kinship with other biological objects. The problems of metrological support of molecular genetic identification by Sanger DNA sequencing using fluorescently labeled reaction terminators and the use of capillary electrophoresis on the example of fish identification are considered. Metrological support of the process includes the use of such definitions as a nucleotide (the nucleic acid monomer), a sequence of nucleotides of nucleic acids. Also, it involves the procedures used in certified methods, a standard sample of a known DNA sequence of an approved type, and a genetic analyzer.Standard Reference Materials of a fragment of human mitochondrial DNA was developed, approved, entered into the Federal registry. The tests of the domestic genetic analyzer Nanophore 05 were carried out. Two measurement procedures “The measurement methodology the partial nucleotide sequence of the control region of mitochondrial DNA of fish (families Acipenseridae and Polyodontidae) by Sanger sequencing with fluorescently-labeled terminators and using capillary electrophoresis” and “The measurement methodology the nucleotide sequence of the 5’ region of the mitochondrial COI gene of water biological resources by Sanger sequencing with fluorescently-labeled terminators and using capillary electrophoresis” were tested and certified. The results are relevant to research in biological sciences, the food industry, biotechnology, and DNA forensics.

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Genetic Analyzer Nanofor 05 as a Measuring Instrument for DNA Sequencing

Cited by 2 publications

References 11 publications

Metrological Support of Nucleic Acid Sequence Analysis

Metrological Support of Nucleic Acid Sequence Analysis

Metrological support of sequence analysis of nucleic acids

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