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Application of the Duplex-Specific Nuclease Preference Method to the Analysis of Single Nucleotide Polymorphisms in Human Genes

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Abstract

A new modification of the single nucleotide polymorphism (SNP) analysis (DSNP, duplex-specific nuclease preference) method using the duplex-specific nuclease from the king crab was proposed. The method was used to study SNPs in the following human genes: kRAS, nRAS, hRAS, and p53, the genes of blood coagulation factor V, methyltetrahydrofolate reductase, prothrombin, and apolipoprotein E and a deletion in the BRCA1 gene. DSNP was shown to be useful for the estimation of the mutant allele content in DNA samples. A system for the simultaneous identification of several adjacent single-nucleotide polymorphisms in the kRAS gene was proposed. The approaches could be used to develop test systems for the detection of SNPs in human genes.

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Abbreviations

DABCYL:

4-((4-(dimethylamino)phenyl)azo)benzoic acid

DSN:

duplex-specific nuclease

DSNP:

duplex-specific nuclease preference method (analysis of oligonucleotide polymorphisms using duplex-specific nuclease)

FAM:

5-carboxyfluorescein

FRET:

resonance transfer of fluorescence energy

MTHFR:

the methyltetrahydrofolate reductase gene

SNP:

single nucleotide polymorphism

TAMPA:

5-carboxytetramethylrhodamine

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Authors and Affiliations

  1. ZAO Evrogen, Moscow, Russia

    I. M. Altshuler

  2. Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow, 117997, Russia

    P. A. Zhulidov, E. A. Bogdanova, N. N. Mudrik & D. A. Shagin

Authors
  1. I. M. Altshuler
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  2. P. A. Zhulidov
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  3. E. A. Bogdanova
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  4. N. N. Mudrik
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  5. D. A. Shagin
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Correspondence to D. A. Shagin.

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__________

Translated from Bioorganicheskaya Khimiya, Vol. 31, No. 6, 2005, pp. 627–636.

Original Russian Text Copyright © 2005 by Altshuler, Zhulidov, Bogdanova, Mudrik, Shagin.

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Altshuler, I.M., Zhulidov, P.A., Bogdanova, E.A. et al. Application of the Duplex-Specific Nuclease Preference Method to the Analysis of Single Nucleotide Polymorphisms in Human Genes. Russ J Bioorg Chem 31, 567–575 (2005). https://doi.org/10.1007/s11171-005-0078-5

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  • DOI: https://doi.org/10.1007/s11171-005-0078-5

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