title: Two-color, 30 second microwave-accelerated Metal-Enhanced Fluorescence DNA assays: a new Rapid Catch and Signal (RCS) technology.
abstract: For analyses of DNA fragment sequences in solution we introduce a 2-color DNA assay, utilizing a combination of the Metal-Enhanced Fluorescence (MEF) effect and microwave-accelerated DNA hybridization. The assay is based on a new "Catch and Signal" technology, i.e. the simultaneous specific recognition of two target DNA sequences in one well by complementary anchor-ssDNAs, attached to silver island films (SiFs). It is shown that fluorescent labels (Alexa 488 and Alexa 594), covalently attached to ssDNA fragments, play the role of biosensor recognition probes, demonstrating strong response upon DNA hybridization, locating fluorophores in close proximity to silver NPs, which is ideal for MEF. Subsequently the emission dramatically increases, while the excited state lifetime decreases. It is also shown that 30s microwave irradiation of wells, containing DNA molecules, considerably (~1000-fold) speeds up the highly selective hybridization of DNA fragments at ambient temperature. The 2-color "Catch and Signal" DNA assay platform can radically expedite quantitative analysis of genome DNA sequences, creating a simple and fast bio-medical platform for nucleic acid analysis.
authors.affiliation: Institute of Fluorescence, University of Maryland Baltimore County, 701 East Pratt Street, Baltimore, MD 21202, USA.
authors.forename: Anatoliy I
authors.initials: AI
authors.lastname: Dragan
authors.forename: Karina
authors.initials: K
authors.lastname: Golberg
authors.forename: Amit
authors.initials: A
authors.lastname: Elbaz
authors.forename: Robert
authors.initials: R
authors.lastname: Marks
authors.forename: Yongxia
authors.initials: Y
authors.lastname: Zhang
authors.forename: Chris D
authors.initials: CD
authors.lastname: Geddes
mesh_terms: D000818:Animals; D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D015342:DNA Probes / Q000235:genetics; D005453:Fluorescence; D005456:Fluorescent Dyes; D006834:Hydrazines; D053768:Metal Nanoparticles; D008872:Microwaves; D009693:Nucleic Acid Hybridization; D009930:Organic Chemicals; D017422:Sequence Analysis, DNA / Q000379:methods* / Q000706:statistics & numerical data; D012834:Silver; D013050:Spectrometry, Fluorescence
chemical_list: C417561:Alexa 488 hydrazide; C417664:Alexa594; D015342:DNA Probes; D005456:Fluorescent Dyes; D006834:Hydrazines; D009930:Organic Chemicals; D012834:Silver