Design and applications of a novel type of Hairpin probes: Adressable Bipartite Molecular Hook (ABMH)

摘要

Dual labelled hairpin probes recognize target sequences with a greater degree of specificity than their linear counterpart. Due to their pre-determined secondary structure, they assume a hairpin loop-stem conformation in the absence of a target, leading to quenching of the fluorescence signal. In the presence of a target nucleic acid, the stem dissociates and separation of the fluorophore from the quencher occurs, allowing emission of fluorescence. The use of molecular beacons (MB) and tripartite molecular beacons (TMB) immobilised on solid supports have simplified microarray protocols. However, the current technologies have problems especially high background and difficulty in preserving a functional structure, extensive loss in signal and specificity upon spotting. We have developed a probe that, for the fist time, displays the solution's characteristics (specificity, sensitivity and discrimination (1 central mismatch)) of MB when ABMH are immobilised on a solid support. We will here present this novel structure with regards to their design and properties and compare these to the current technologies (especially MB).