Polyacrylamide Gel Film Immobilized Molecular Beacon Array for Single Nucleotide Mismatch Detection
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.
Document Type: Research Article
Publication date: April 1, 2005
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