Issue 13, 2018

Simultaneous multiple single nucleotide polymorphism detection based on click chemistry combined with DNA-encoded probes

Abstract

Single nucleotide polymorphisms (SNPs) are emerging as important biomarkers for disease diagnosis, prognostics and disease pathogenesis. As one type of disease is always connected to several SNP sites, there is great demand for a reliable multiple SNP detection method. Herein, we mimicked a ligation reaction based on DNA ligase and originally utilized an enzyme-free DNA template-directed click reaction for SNP detection. With 5′-alkyne and 3′-azide groups labelled on two oligonucleotide probes, the target DNA-directed Cu(I)-catalyzed alkyne–azide cycloaddition (CuAAC) click reaction produced a new DNA strand with a triazole backbone, as a mimic of a DNA phosphodiester linkage. Trace amounts of the target (as low as 25 fmol in 50 μL) could be sensitively detected using capillary gel electrophoresis with laser-induced fluorescence (CGE-LIF). Meanwhile, SNP caused an obvious difference in the efficiency of the click reaction, and 0.5% SNP could be easily detected. More importantly, multiplexed SNP detection in a one tube reaction was successfully achieved only by encoding different lengths of the DNA probes for the different SNP sites.

Graphical abstract: Simultaneous multiple single nucleotide polymorphism detection based on click chemistry combined with DNA-encoded probes

Supplementary files

Article information

Article type
Edge Article
Submitted
19 Jan 2018
Accepted
21 Feb 2018
First published
22 Feb 2018
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY license

Chem. Sci., 2018,9, 3335-3340

Simultaneous multiple single nucleotide polymorphism detection based on click chemistry combined with DNA-encoded probes

Q. Zhou, F. Yuan, X. Zhang, Y. Zhou and X. Zhang, Chem. Sci., 2018, 9, 3335 DOI: 10.1039/C8SC00307F

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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