Detection of DNA hybridization via fluorescent polymer superquenching

被引：128

作者：

Kushon, SA ^{[1
]}

Ley, KD ^{[1
]}

Bradford, K ^{[1
]}

Jones, RM ^{[1
]}

McBranch, D ^{[1
]}

Whitten, D ^{[1
]}

机构：

[1] QTL Biosyst, LLC, Santa Fe, NM 87507 USA

来源：

LANGMUIR | 2002年 / 18卷 / 20期

关键词：

D O I：

10.1021/la026211u

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

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.

引用

页码：7245 / 7249

页数：5

共 28 条

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