Multicolor molecular beacons for allele discrimination.
Molecular beacons are hairpin-shaped oligonucleotide probes that report the presence of specific nucleic acids in homogenous solutions. When they bind to their targets they undergo a conformational reorganization that restores the fluorescence of an internally quenched fluorophore. We found that their hairpin conformation enables the use of a wide variety of differently colored fluorophores. Using several molecular beacons, each designed to recognize a different target and each labeled with a different fluorophore, we demonstrate that multiple targets can be distinguished in the same solution, even if they differ from one another by as little as a single nucleotide. A comparison of "hairpin probes" with corresponding "linear probes" confirms that the presence of the hairpin stem in molecular beacons significantly enhances their specificity.
Tyagi
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S
Department of Molecular Genetics, Public Health Research Institute, New York, NY 10016, USA. sanjay@phri.nyu.edu
Bratu
D P
DP
Kramer
F R
FR
D000483:Alleles; D017931:DNA Primers; D005456:Fluorescent Dyes; D009154:Mutation / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D013050:Spectrometry, Fluorescence; D004124:p-Dimethylaminoazobenzene / Q000031:analogs & derivatives
D017931:DNA Primers; D005456:Fluorescent Dyes; D015345:Oligonucleotide Probes; C063127:4-(4-dimethylaminophenylazo)benzoic acid; D004124:p-Dimethylaminoazobenzene