title: Label-free hybridization detection of a single nucleotide mismatch by immobilization of molecular beacons on an agarose film.
abstract: We developed a new 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.
authors.affiliation: Chien-Shiung Wu Laboratory, Department of Biomedical Engineering, Southeast University, Nanjing, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.forename: Jiong
authors.initials: J
authors.lastname: Li
authors.forename: Heping
authors.initials: H
authors.lastname: Liu
authors.forename: Quanjun
authors.initials: Q
authors.lastname: Liu
authors.forename: Qian
authors.initials: Q
authors.lastname: Mei
authors.forename: Yijin
authors.initials: Y
authors.lastname: Wang
authors.forename: Jijun
authors.initials: J
authors.lastname: Zhu
authors.forename: Nongyue
authors.initials: N
authors.lastname: He
authors.forename: Zuhong
authors.initials: Z
authors.lastname: Lu
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000032:analysis; D005453:Fluorescence; D005976:Glutaral / Q000737:chemistry; D020180:Heteroduplex Analysis / Q000379:methods*; D007700:Kinetics; D009692:Nucleic Acid Heteroduplexes / Q000032:analysis*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015345:Oligonucleotide Probes; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D012685:Sepharose / Q000737:chemistry*
chemical_list: D009692:Nucleic Acid Heteroduplexes; D015345:Oligonucleotide Probes; D004247:DNA; D012685:Sepharose; D005976:Glutaral