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Mikrochim Acta ; 185(6): 299, 2018 05 13.
Article in English | MEDLINE | ID: mdl-29754198

ABSTRACT

The authors describe a novel assay for the detection of methylated DNA site. Rolling circle amplification and CdSe/ZnS quantum dots with high fluorescence efficiency are applied in this method. The CdSe/ZnS quantum dots act as electron donors, and hemin and oxygen (derived from hydrogen peroxide act as acceptors in photoinduced electron transfer. The assay, best performed at excitation/emission peaks of 450/620 nm, is sensitive and specific. Fluorometric response is linear in the 1 pM to 100 nM DNA concentration range, and the lowest detectable concentration of methylated DNA is 142 fM (S/N = 3). The method is capable of recognizing 0.01% methylated DNA in a mixture of methylated/unmethylated DNA. Graphical abstract A novel method for methylated sites detection in DNA is established. Rolling circle amplification and photoinduced electron transfer. CdSe/ZnS quantum dots with high fluorescence efficiency act as the electron donor, while G-quadruplex/hemin and hydrogen peroxide derived oxygen act as electron acceptor. It presents a linear response towards 1 pM to 100 nM methylated DNA with a correlation coefficient of 0.9968, and the lowest detectable concentration of methylated DNA was 142 fM, with selectivity significantly superior to other methods.


Subject(s)
DNA Methylation , DNA/chemistry , DNA/genetics , Fluorometry/methods , Limit of Detection , Nucleic Acid Amplification Techniques , Photochemical Processes , Base Sequence , Cadmium Compounds/chemistry , Electron Transport , Quantum Dots/chemistry , Selenium Compounds/chemistry , Sulfides/chemistry , Zinc Compounds/chemistry
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