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The use of duplex-specific crab nuclease for rapid analysis of single-nucleotide polymorphisms and the detection of DNA targets in complex PCR products

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Abstract

A rapid method for single-nucleotide polymorphism analysis and the detection of target clones after cloning of complex PCR products based on the use of duplex-specific crab nuclease and a universal fluorescent probe has been developed. The method is an alternative to the labor-intensive procedures of clone screening employing radioactively labeled probes, gel-based restriction analysis, and costly sequencing. The efficiency of the novel method has been demonstrated in a range of model systems.

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Abbreviations

DSN:

duplex-specific nuclease

ds:

double-stranded

ss:

single-stranded

SNP:

single-nucleotide polymorphism

SP:

specific primer

Dabcyl:

4-(4′-dimethylaminophenylazo) benzoic acid

Fam:

6-carboxyfluorescein

FRET:

fluorescence resonance energy transfer

Tamra:

tetramethylrhodamin

TCR:

T-cell receptor

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Authors and Affiliations

  1. ZAO Evrogen, Moscow, Russia

    I. A. Shagina, I. M. Altshuler & D. A. Shagin

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

    E. A. Bogdanova, S. A. Luk’yanov & D. A. Shagin

Authors
  1. I. A. Shagina
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  2. E. A. Bogdanova
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  3. I. M. Altshuler
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  4. S. A. Luk’yanov
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  5. D. A. Shagin
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Corresponding author

Correspondence to D. A. Shagin.

Additional information

Original Russian Text © I.A. Shagina, E.A. Bogdanova, I.M. Altshuler, S.A. Luk’yanov, D.A. Shagin, 2011, published in Bioorganicheskaya Khimiya, 2011, Vol. 37, No. 4, pp. 522–529.

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Shagina, I.A., Bogdanova, E.A., Altshuler, I.M. et al. The use of duplex-specific crab nuclease for rapid analysis of single-nucleotide polymorphisms and the detection of DNA targets in complex PCR products. Russ J Bioorg Chem 37, 464–471 (2011). https://doi.org/10.1134/S1068162011040121

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  • DOI: https://doi.org/10.1134/S1068162011040121

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