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Fluorescence Detection of Single-nucleotide Polymorphisms Using a Thymidine-based Molecular Beacon by Chi-Wei Liu; Yang-Wei Lin; Chih-Ching Huang; Huan-Tsung Chang is a Engineering article available to read on EtoBox.

What is Fluorescence Detection of Single-nucleotide Polymorphisms Using a Thymidine-based Molecular Beacon about?

We have developed a universal molecular beacon (T(7)-MB-T(7)) 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 (DNA(pm)) and single-base mismatched (DNA(mm)) DNA strands are stronger and weaker, respectively, than those with Hg(2+) ions. As a result, the fluorescence of a solution containing T(7)-MB-T(7), DNA(pm), EthBr, and Hg(2+) is higher than that of a corresponding solution containing T(7)-MB-T(7), DNA(mm), EthBr, and Hg(2+), because the former has a greater number of intercalation sites for EthBr. Under the optimal conditions (100 nM T(7)-MB-T(7), 20 mM NaCl, 5.0 microM Hg(2+), and 300 nM EthBr in 5.0 mM Tris-HCl solution, pH 7.4), the plot of the fluorescence intensity against the concentration of DNA(pm) was linear over the range 5.0-100 nM (R(2)=0.98). A similar probe, T(7)-MB(t)-T(7), is sensitive and selective for the detection of a gene associated wit

Who reads Fluorescence Detection of Single-nucleotide Polymorphisms Using a Thymidine-based Molecular Beacon?

Author

Chi-Wei Liu; Yang-Wei Lin; Chih-Ching Huang; Huan-Tsung Chang

Publisher

Elsevier Science; Elsevier ; Elsevier Ltd.; Elsevier BV (ISSN 0956-5663)

Published

2009

Language

EN

Field

Engineering (Physical Sciences)