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Microchimica Acta

, 186:716 | Cite as

Graphene oxide and enzyme-assisted dual-cycling amplification method for sensitive fluorometric determination of DNA

  • Idorenyin Iwe
  • Zhigang LiEmail author
  • Jiahao HuangEmail author
Original Paper
  • 132 Downloads

Abstract

A fluorometric method is described for the determination of DNA. It involves the use of graphene oxide (GO), exonuclease III (Exo III), and two specially designed fluorophore-labeled hairpin probes (HP1 and HP2). Different from other GO-based DNA assays, the method takes advantage of the distinct binding abilities of GO with hairpin DNA probes and single nucleotides. GO serves as a strong quencher for fluorescent labels to ensure a very low background signal. Two reaction cycles mediated by Exo III are employed to enhance the signals. The combination of GO-induced quenching and Exo III-mediated dual regeneration of analytes leads to a detection limit as low as 1 pM for the model analyte human hemochromatosis protein (HFE) gene. The method is also applicable for the determination of HFE gene spiked into fetal bovine serum.

Graphical abstract

Schematic representation of a GO-based, Exo III-assisted method for dual-signal amplified detection of DNA, for which human haemochromatosis protein (HFE) gene is designed as the model target. The assay involves graphene oxide (GO), exonuclease (Exo III), and two specially designed, fluorophore-labelled hairpin probes (HP1 and HP2).

Keywords

DNA sensor Fluorescence Exonuclease III Nanomaterial Human hemochromatosis protein gene Fetal bovine serum 

Notes

Acknowledgements

This work was partially supported by the Research Grants Council of the Hong Kong Special Administrative Region under Grant No. 16228216 and the National Natural Science Foundation of China under Grant No. 21705076. We would also like to acknowledge the supply of graphene oxide from World Linkage Holdings Limited (Hong Kong).

Compliance with ethical standards

Conflict of interest

The author(s) declare that they have no conflict of interest.

Supplementary material

604_2019_3825_MOESM1_ESM.docx (407 kb)
ESM 1 (DOCX 406 kb)

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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Mechanical and Aerospace EngineeringThe Hong Kong University of Science and TechnologyKowloonChina
  2. 2.Guangdong Provincial Key Laboratory of Construction and Detection in Tissue Engineering, Biomaterials Research Center, School of Biomedical EngineeringSouthern Medical UniversityGuangzhouChina

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