Label-free hybridization detection of a single nucleotide mismatch by immobilization of molecular beacons on an agarose film

We developed a now technique to Immobilize a set of molecular beacons on an agarose film-coated slide and found that It has the ability to Identify a single nucleotide difference in label-free DNA targets. The annealing properties, specificity and hybridization dynamics of the present technique were compared with those of the conventional technique that directly Immobilizes molecular beacons on a planar glass slide. It Is demonstrated that the molecular beacon array on an agarose film has high quench efficiency, an excellent discrimination ratio for single nucleotide mismatches and a short detection time. We hypothesize that such a low fluorescence background and high specificity molecular beacon array will find practical applications in label-free, high-throughput mutation analysis and disease diagnosis.