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

  • I. A. Shagina
  • E. A. Bogdanova
  • I. M. Altshuler
  • S. A. Luk’yanov
  • D. A. Shagin
Article
  • 69 Downloads

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.

Keywords

duplex-specific crab nuclease polymerase chain reaction single-nucleotide polymorphisms 

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

Copyright information

© Pleiades Publishing, Ltd. 2011

Authors and Affiliations

  • I. A. Shagina
    • 1
  • E. A. Bogdanova
    • 2
  • I. M. Altshuler
    • 1
  • S. A. Luk’yanov
    • 2
  • D. A. Shagin
    • 1
    • 2
  1. 1.ZAO EvrogenMoscowRussia
  2. 2.Shemyakin-Ovchinnikov Institute of Bioorganic ChemistryRussian Academy of SciencesMoscowRussia

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