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Spectroscopic Study of CpG Alternating DNA-Methylene Blue Interaction for Methylation Detection

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Abstract

Recognition of methylated DNA sites would be useful strategy due to the important roles of methylation in disease occurrence and developmental processes. The interaction of CpG rich methylated and unmethylated DNA hybrid with methylene blue (MB) as an optical probe has been investigated by absorption, emission, circular dichorism and fluorescence anisotropy analysis. Titration of MB with both sequences caused a hypsochromism and decreased the absorption of MB that indicating an intercalative mode of interaction. The experimental results revealed that MB as the optical indicator could distinguish between the methylated and unmethylated DNA sequences. Under optimum conditions, upon the addition of methylated dsDNA, the fluorescence intensity increased in linear range from 1.0 × 10−9 to 1.0 × 10−6 M with detection limit of 7.2 × 10−10 M and on the other hand, the intensity of MB showed no change with addition of unmethylated dsDNA.

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Acknowledgments

The authors thank the research Council of University of Tehran for financial support of this work.

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Authors and Affiliations

  1. Department of Life Science Engineering, Faculty of New Sciences & Technologies, University of Tehran, Tehran, Iran

    Morteza Hosseini & Fereshteh Khaki

  2. Department of Biotechnology, Payame Noor University, Tehran, Iran

    Mehdi Dadmehr

  3. Center of Excellence in Electrochemistry, Faculty of Chemistry, University of Tehran, Tehran, Iran

    Mohammad Reza Ganjali

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  1. Morteza Hosseini
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  2. Fereshteh Khaki
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  3. Mehdi Dadmehr
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  4. Mohammad Reza Ganjali
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Correspondence to Morteza Hosseini.

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Hosseini, M., Khaki, F., Dadmehr, M. et al. Spectroscopic Study of CpG Alternating DNA-Methylene Blue Interaction for Methylation Detection. J Fluoresc 26, 1123–1129 (2016). https://doi.org/10.1007/s10895-016-1804-5

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  • DOI: https://doi.org/10.1007/s10895-016-1804-5

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