title: Fluorometric Detection of MicroRNA Using Isothermal Gene Amplification and Graphene Oxide.
abstract: We have developed a facile fluorometric system for the detection of microRNA (miRNA), using rolling circle amplification (RCA), graphene oxide (GO), and fluorescently labeled peptide nucleic acid (F-PNA). The padlock probe DNA complementary to a target miRNA was selectively ligated to form circular DNA that was then used as the template for RCA. F-PNAs complementary to the target miRNA were annealed to multiple sites of the isothermally amplified single-stranded RCA product (RCAP) containing multiple target miRNA sequences. This F-PNA/RCAP duplex is less adsorbed onto the GO monolayer, thus attenuating the quenching of F-PNA fluorescence by GO. In the absence of target miRNA (and hence the absence of RCA and duplex formation), the free F-PNA is completely adsorbed onto the GO monolayer and fluorescence quenching ensues. Thus, GO-based fluorescence detection coupled with isothermal gene amplification would be a simple and convenient method for the quantitative detection of miRNA.
authors.affiliation: Department of Bioscience and Biotechnology, Konkuk University , Seoul 143-701, Korea.
authors.forename: Chaesun
authors.initials: C
authors.lastname: Hong
authors.affiliation: Department of Bioscience and Biotechnology, Konkuk University , Seoul 143-701, Korea.
authors.forename: Ahruem
authors.initials: A
authors.lastname: Baek
authors.forename: Sang Soo
authors.initials: SS
authors.lastname: Hah
authors.forename: Woong
authors.initials: W
authors.lastname: Jung
authors.affiliation: Department of Bioscience and Biotechnology, Konkuk University , Seoul 143-701, Korea.
authors.forename: Dong-Eun
authors.initials: DE
authors.lastname: Kim
mesh_terms: D005470:Fluorometry / Q000379:methods*; D005784:Gene Amplification; D006108:Graphite / Q000737:chemistry*; D035683:MicroRNAs / Q000737:chemistry*; D010087:Oxides / Q000737:chemistry
chemical_list: D035683:MicroRNAs; D010087:Oxides; D006108:Graphite