title: An exonuclease III and graphene oxide-aided assay for DNA detection.
abstract: We have developed a novel DNA assay based on exonuclease III (ExoIII)-induced target recycling and the fluorescence quenching ability of graphene oxide (GO). This assay consists of a linear DNA probe labeled with a fluorophore in the middle. Introduction of target sequence induces the exonuclease III catalyzed probe digestion and generation of single nucleotides. After each cycle of digestion, the target is recycled to realize the amplification. Finally, graphene oxide is added to quench the remaining probes and the signal from the resulting fluorophore labeled single nucleotides is detected. With this approach, a sub-picomolar detection limit can be achieved within 40 min at 37°C. The method was successfully applied to multicolor DNA detection and the analysis of telomerase activity in extracts from cancer cells.
authors.affiliation: Center for Research at Bio/nano Interface, Department of Chemistry and Department of Physiology and Functional Genomics, Shands Cancer Center, University of Florida, Gainesville, FL 32611-7200, USA.
authors.forename: Lu
authors.initials: L
authors.lastname: Peng
authors.affiliation: Center for Research at Bio/nano Interface, Department of Chemistry and Department of Physiology and Functional Genomics, Shands Cancer Center, University of Florida, Gainesville, FL 32611-7200, USA.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Zhu
authors.affiliation: Center for Research at Bio/nano Interface, Department of Chemistry and Department of Physiology and Functional Genomics, Shands Cancer Center, University of Florida, Gainesville, FL 32611-7200, USA.
authors.forename: Yan
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Center for Research at Bio/nano Interface, Department of Chemistry and Department of Physiology and Functional Genomics, Shands Cancer Center, University of Florida, Gainesville, FL 32611-7200, USA.
authors.forename: Da
authors.initials: D
authors.lastname: Han
authors.affiliation: Center for Research at Bio/nano Interface, Department of Chemistry and Department of Physiology and Functional Genomics, Shands Cancer Center, University of Florida, Gainesville, FL 32611-7200, USA. Electronic address: tan@chem.ufl.edu.
authors.forename: Weihong
authors.initials: W
authors.lastname: Tan
mesh_terms: D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods* / Q000706:statistics & numerical data; D004247:DNA / Q000032:analysis* / Q000235:genetics; D015342:DNA Probes; D005090:Exodeoxyribonucleases; D006108:Graphite; D006367:HeLa Cells; D006801:Humans; D057230:Limit of Detection; D021141:Nucleic Acid Amplification Techniques; D012680:Sensitivity and Specificity; D019098:Telomerase / Q000032:analysis
chemical_list: D015342:DNA Probes; D006108:Graphite; D004247:DNA; D019098:Telomerase; D005090:Exodeoxyribonucleases; C020133:exodeoxyribonuclease III