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- W2014920121 abstract "We have developed a universal molecular beacon (T7-MB-T7) for the detection of single-nucleotide polymorphisms (SNPs). The beacon, which contains a 19-mer loop and a stem comprising a pair of seven thymidine (T) bases, forms double-stranded structures with target DNA molecules, leading to increases in the fluorescence of ethidium bromide (EthBr) as a result of intercalation. The interactions of the beacon with perfectly matched (DNApm) and single-base mismatched (DNAmm) DNA strands are stronger and weaker, respectively, than those with Hg2+ ions. As a result, the fluorescence of a solution containing T7-MB-T7, DNApm, EthBr, and Hg2+ is higher than that of a corresponding solution containing T7-MB-T7, DNAmm, EthBr, and Hg2+, because the former has a greater number of intercalation sites for EthBr. Under the optimal conditions (100 nM T7-MB-T7, 20 mM NaCl, 5.0 μM Hg2+, and 300 nM EthBr in 5.0 mM Tris–HCl solution, pH 7.4), the plot of the fluorescence intensity against the concentration of DNApm was linear over the range 5.0–100 nM (R2 = 0.98). A similar probe, T7-MBt-T7, is sensitive and selective for the detection of a gene associated with hereditary tyrosinemia type I. Relative to conventional MBs, our new probe offers the advantages of higher selectivity toward DNA, less nonspecific binding toward single-stranded-DNA-binding protein, greater resistance to nuclease digestion, and low cost; therefore, we suspect that this system holds great potential for practical studies of SNPs." @default.
- W2014920121 created "2016-06-24" @default.
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- W2014920121 date "2009-04-01" @default.
- W2014920121 modified "2023-09-25" @default.
- W2014920121 title "Fluorescence detection of single-nucleotide polymorphisms using a thymidine-based molecular beacon" @default.
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- W2014920121 doi "https://doi.org/10.1016/j.bios.2009.01.003" @default.
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