Screening unlabeled DNA targets with randomly ordered fiber-optic gene arrays.
We have developed a randomly ordered fiber-optic gene array for rapid, parallel detection of unlabeled DNA targets with surface immobilized molecular beacons (MB) that undergo a conformational change accompanied by a fluorescence change in the presence of a complementary DNA target. Microarrays are prepared by randomly distributing MB-functionalized 3-microm diameter microspheres in an array of wells etched in a 500-microm diameter optical imaging fiber. Using several MBs, each designed to recognize a different target, we demonstrate the selective detection of genomic cystic fibrosis related targets. Positional registration and fluorescence response monitoring of the microspheres was performed using an optical encoding scheme and an imaging fluorescence microscope system.
Steemers
F J
FJ
The Max Tishler Laboratory for Organic Chemistry, Department of Chemistry, Tufts University, Medford, MA.
Ferguson
J A
JA
Walt
D R
DR
D001483:Base Sequence; D001665:Binding Sites; D001710:Biotin / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D004247:DNA / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D005336:Fiber Optic Technology; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D008856:Microscopy, Fluorescence; D008863:Microspheres; D015336:Molecular Probe Techniques; D009154:Mutation / Q000235:genetics; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015345:Oligonucleotide Probes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D055100:Optical Fibers; D016133:Polymerase Chain Reaction; D011897:Random Allocation; D012680:Sensitivity and Specificity; D013997:Time Factors
C505032:CFTR protein, human; D005456:Fluorescent Dyes; D015345:Oligonucleotide Probes; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D001710:Biotin; D004247:DNA