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Polyacrylamide Gel Film Immobilized Molecular Beacon Array for Single Nucleotide Mismatch Detection

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Abstract:

We reported polyacrylamide gel immobilized molecular beacon array for single nucleotide mismatch detection in this paper. Molecular beacons are oligonucleotide probes fluorescing upon hybridization to their complementary DNA/RNA targets with excellent sensitivity and high selectivity. The specially designed molecular beacon for immobilization contains a 15 base loop sequence with a 5 base pair stem, a polyT (20 bases) spacer, a 5′-end amino group for immobilization, a fluorescein in the middle of the sequence as the fluorophore, and a 3′-end DABCYL as the quencher. Between the 5′-end amino group and the stem, the polyT is used to minimize disability caused by 5′-end immobilization. The molecular beacon microarray was fabricated by a pin-based spotting robot and the hybridization was investigated by confocal microscope. A real-time hybridization process at room temperature was registered every minute for 20 min after the target solution was pumped into the hybridization cell. The result indicates that a polyacrylamide film coated glass slide provides an ideal solution-like environment for molecular beacon probes. The potential applications of this kind of molecular beacon array are mutation detection, disease mechanisms, disease diagnostics, etc. in a parallel, cost saving, and label-free detection way.

Keywords: MICROARRAY; MOLECULAR BEACON; MUTATION DETECTION; POLYACRYLAMIDE GEL; SINGLE NUCLEOTIDE MISMATCH DETECTION

Document Type: Research Article

DOI: http://dx.doi.org/10.1166/jnn.2005.086

Publication date: April 1, 2005

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  • Journal for Nanoscience and Nanotechnology (JNN) is an international and multidisciplinary peer-reviewed journal with a wide-ranging coverage, consolidating research activities in all areas of nanoscience and nanotechnology into a single and unique reference source. JNN is the first cross-disciplinary journal to publish original full research articles, rapid communications of important new scientific and technological findings, timely state-of-the-art reviews with author's photo and short biography, and current research news encompassing the fundamental and applied research in all disciplines of science, engineering and medicine.
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