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Issue 7, 2008
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Analysis of DNA and single-base mutations using magnetic particles for purification, amplification and DNAzyme detection

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Abstract

The amplified detection of DNA or of single-base mismatches in DNA is achieved by the use of nucleic acid-functionalized magnetic particles that separate the recognition duplexes and, upon amplification, yield chemiluminescence-generating DNAzymes as reporter units. The analysis of M13 phage ssDNA is achieved by the hybridization of the analyte to capture nucleic acid-functionalized magnetic particles followed by the binding of a DNA machine unit to the analyte domain. The magnetic separation of the multi-component-functionalized magnetic particles, followed by their reaction with polymerase, dNTPs, and the nicking enzyme (Nb.BbvCI) activate the autonomous synthesis of the horseradish peroxidase-mimicking DNAzyme that acts as chemiluminescent reporter. The single-base mutation in DNA is achieved by coupling of the DNA machine to the mutant DNA/capture nucleic acid-functionalized magnetic particles hybrid structure. The activation of the polymerization/nicking cycles yield the chemiluminescent reporting DNAzyme. The magnetic separation of the DNA recognition hybrids improves the signal-to-noise ratio of the analytical protocol as compared to related DNAzyme synthesizing schemes.

Graphical abstract: Analysis of DNA and single-base mutations using magnetic particles for purification, amplification and DNAzyme detection

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Article information


Submitted
05 Feb 2008
Accepted
17 Mar 2008
First published
23 Apr 2008

Analyst, 2008,133, 923-927
Article type
Paper

Analysis of DNA and single-base mutations using magnetic particles for purification, amplification and DNAzyme detection

I. Willner, Z. Cheglakov, Y. Weizmann and E. Sharon, Analyst, 2008, 133, 923
DOI: 10.1039/B802015A

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