title: Effect of salts, solvents and buffer on miRNA detection using DNA silver nanocluster (DNA/AgNCs) probes.
abstract: MicroRNAs (miRNAs) are small regulatory RNAs (size ~21 nt to ~25 nt) which regulate a variety of important cellular events in plants, animals and single cell eukaryotes. Especially because of their use in diagnostics of human diseases, efforts have been directed towards the invention of a rapid, simple and sequence selective detection method for miRNAs. Recently, we reported an innovative method for the determination of miRNA levels using the red fluorescent properties of DNA/silver nanoclusters (DNA/AgNCs). Our method is based on monitoring the emission drop of a DNA/AgNCs probe in the presence of its specific target miRNA. Accordingly, the accuracy and efficiency of the method relies on the sensitivity of hybridization between the probe and target. To gain specific and robust hybridization between probe and target, we investigated a range of diverse salts, organic solvents, and buffer to optimize target sensing conditions. Under the newly adjusted conditions, the target sensitivity and the formation of emissive DNA/AgNCs probes were significantly improved. Also, fortification of the Tris-acetate buffer with inorganic salts or organic solvents improved the sensitivity of the DNA/AgNC probes. On the basis of these optimizations, the versatility of the DNA/AgNCs-based miRNA detection method can be expanded.
authors.affiliation: UNIK Center for Synthetic Biology/Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 4, DK-1871 Frederiksberg C, Copenhagen, Denmark.
authors.forename: Pratik
authors.initials: P
authors.lastname: Shah
authors.forename: Seok Keun
authors.initials: SK
authors.lastname: Cho
authors.forename: Peter Waaben
authors.initials: PW
authors.lastname: Thulstrup
authors.forename: Yong-Joo
authors.initials: YJ
authors.lastname: Bhang
authors.forename: Jong Cheol
authors.initials: JC
authors.lastname: Ahn
authors.forename: Suk Won
authors.initials: SW
authors.lastname: Choi
authors.forename: Andreas
authors.initials: A
authors.lastname: Rørvig-Lund
authors.forename: Seong Wook
authors.initials: SW
authors.lastname: Yang
mesh_terms: D002021:Buffers; D004247:DNA / Q000737:chemistry*; D015342:DNA Probes / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry; D053768:Metal Nanoparticles / Q000737:chemistry*; D035683:MicroRNAs / Q000032:analysis*; D009693:Nucleic Acid Hybridization; D012492:Salts / Q000737:chemistry*; D012834:Silver / Q000737:chemistry*; D012997:Solvents / Q000737:chemistry*; D013050:Spectrometry, Fluorescence
chemical_list: D002021:Buffers; D015342:DNA Probes; D005456:Fluorescent Dyes; D035683:MicroRNAs; D012492:Salts; D012997:Solvents; D012834:Silver; D004247:DNA