Issue 8, 2011
From the journal:

Analyst

Visual SNP genotyping using asymmetric PCR and split DNA enzymes†

Jia Ling Neo,a   Kanglie Darius Awb  and  Mahesh Uttamchandani*ac  

* Corresponding authors

a Defence Medical and Environmental Research Institute, DSO National Laboratories, 27 Medical Drive, Singapore
E-mail: mahesh@dso.org.sg
Fax: +65 6485 7033

b School of Chemical and Biomedical Engineering, Nanyang Technological University, N1.2-B3-13, 62 Nanyang Drive, Singapore

c Departments of Chemistry and Biological Sciences, National University of Singapore, 3 Science Drive 3, Singapore

Abstract

Harnessing and applying genomic technologies in resource limited environments demand a next generation of platforms, which are convenient, quick and easy to apply. We describe here a visual colour change assay that can be applied to SNP genotyping, which unlike traditional methods, does not adopt complicated procedures or expensive instrumentation, desirable features in bringing genetic capabilities outside the laboratory. Our strategy involved a two-step method that first enriched target genomic regions using asymmetric PCR, followed by direct in situ application of split DNA probes. In the presence of target sequences that perfectly matched the complementary probes, the split probes reassembled active DNAzymes, which catalysed a colour change reaction. A single-base mismatch (indicative of a polymorphism) prevented this reassembly and colour change, providing the means for accurate SNP calling. This is the first report, to our knowledge, that demonstrates successful visual SNP genotyping of actual human DNA samples using DNAzymes.

Graphical abstract: Visual SNP genotyping using asymmetric PCR and split DNA enzymes

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Article information

DOI
https://doi.org/10.1039/C0AN00838A
Article type
Communication
Submitted
29 Oct 2010
Accepted
20 Feb 2011
First published
07 Mar 2011

Analyst, 2011,136, 1569-1572

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Visual SNP genotyping using asymmetric PCR and split DNA enzymes

J. L. Neo, K. D. Aw and M. Uttamchandani, Analyst, 2011, 136, 1569 DOI: 10.1039/C0AN00838A

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