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Engineering a DNA-cleaving DNAzyme and PCR into a simple sensor for zinc ion detection

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Abstract

The development of a simple sensor (9NL27-Zn) based on DNAzyme and PCR and aimed at the detection of low concentrations of zinc (II) ions is described. A specific Zn(II)-dependent DNAzyme (9NL27) with DNA-cleaving activity was employed. In the presence of zinc (II), the DNAzyme hydrolyzed DNA substrate into two pieces (5′ and 3′ fragments), forming 3′-terminal hydroxyl in the 5′ fragment and 5′-phosphate in the 3′ fragments. Subsequently, the 5′ fragment left the DNAzyme and bound a short DNA template. The 5′ fragment was used as a primer and extended a single-stranded full-length template by Taq polymerase. Finally, this full-length template was amplified by PCR. The amplified products had a quantitative relationship with Zn(II) concentration. Under our experimental conditions, the DNA sensor showed sensitivity (10 nM) and high specificity for zinc ion detection. After improvement of the DNA sensor, the detection limit can reach 1 nM. The simple DNA sensor may become a DNA model for the detection of trace amounts of other targets.

The general principle of a 9NL27-Zn sensor by the combination of a Zn2+-dependent DNAzyme and PCR. The red arrowhead indicates the cleavage site of DNA substrate

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Acknowledgments

This research was supported by the Fundamental Research Funds for Jilin University (200903096) and National Natural Science Foundation of China (31100573).

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  1. Jiacui Xu

    Present address: Department of Biochemistry, University of Wisconsin, Madison, WI, 53706, USA

Authors and Affiliations

  1. Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, College of Life Science, Jilin University, Qianjin Street 2699, 130012, Changchun, China

    Jiacui Xu, Yanhong Sun, Yongjie Sheng, Yanqun Fei, Jin Zhang & Dazhi Jiang

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  1. Jiacui Xu
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  2. Yanhong Sun
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  3. Yongjie Sheng
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  4. Yanqun Fei
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  5. Jin Zhang
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Correspondence to Jin Zhang or Dazhi Jiang.

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Xu, J., Sun, Y., Sheng, Y. et al. Engineering a DNA-cleaving DNAzyme and PCR into a simple sensor for zinc ion detection. Anal Bioanal Chem 406, 3025–3029 (2014). https://doi.org/10.1007/s00216-014-7732-9

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  • DOI: https://doi.org/10.1007/s00216-014-7732-9

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