Label-free hybridization detection of a single nucleotide mismatch by immobilization of molecular beacons on an agarose film.
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.
Wang
Hong
H
Chien-Shiung Wu Laboratory, Department of Biomedical Engineering, Southeast University, Nanjing, China.
Li
Jiong
J
Liu
Heping
H
Liu
Quanjun
Q
Mei
Qian
Q
Wang
Yijin
Y
Zhu
Jijun
J
He
Nongyue
N
Lu
Zuhong
Z
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*
D009692:Nucleic Acid Heteroduplexes; D015345:Oligonucleotide Probes; D004247:DNA; D012685:Sepharose; D005976:Glutaral