Detection of DNA Hybridization via Fluorescent Polymer Superquenching

An assay for a target single strand 20-base sequence of DNA coding for the anthrax lethal factor, based on conjugated polymer fluorescence superquenching, is reported. The assay employs a platform in which the receptor (a biotinylated complementary sequence “capture strand”) and polymer (two compone...

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Veröffentlicht in:Langmuir 2002-10, Vol.18 (20), p.7245-7249
Hauptverfasser: Kushon, Stuart A, Ley, Kevin D, Bradford, Kirsten, Jones, Robert M, McBranch, Duncan, Whitten, David
Format: Artikel
Sprache:eng
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Zusammenfassung:An assay for a target single strand 20-base sequence of DNA coding for the anthrax lethal factor, based on conjugated polymer fluorescence superquenching, is reported. The assay employs a platform in which the receptor (a biotinylated complementary sequence “capture strand”) and polymer (two components:  an anionic poly(phenylene ethynylene) (PPE) and a biotinylated −PPE) are co-located on streptavidin-derivatized polystyrene microspheres. A conjugate of the target strand with the energy transfer quencher QSY-7 (DNA-QTL) is used to construct competition assays for the target. A direct competition assay between the target-DNA and DNA-QTL for the microsphere-bound capture is only marginally successful due evidently to greater kinetic affinity of the polymer-capture ensemble for the conjugate. However a sequential addition of target, followed by DNA-QTL affords a quantitative assay for the target by attenuation of PPE fluorescence quenching by the DNA-QTL. Likewise a direct competition in solution between the target and DNA-QTL for the biotinylated capture strand followed by addition of microspheres provides a sensitive and quantitative assay for the target single strand DNA.
ISSN:0743-7463
1520-5827
DOI:10.1021/la026211u