Molecular-beacon-based array for sensitive DNA analysis.
Molecular beacon (MB) DNA probes provide a new way for sensitive label-free DNA/protein detection in homogeneous solution and biosensor development. However, a relatively low fluorescence enhancement after the hybridization of the surface-immobilized MB hinders its effective biotechnological applications. We have designed new molecular beacon probes to enable a larger separation between the surface and the surface-bound MBs. Using these MB probes, we have developed a DNA array on avidin-coated cover slips and have improved analytical sensitivity. A home-built wide-field optical setup was used for imaging the array. Our results show that linker length, pH, and ionic strength have obvious effects on the performance of the surface-bound MBs. The fluorescence enhancement of the new MBs after hybridization has been increased from 2 to 5.5. The MB-based DNA array could be used for DNA detection with high sensitivity, enabling simultaneous multiple-target bioanalysis in a variety of biotechnological applications.
Yao
Gang
G
Center for Research at the Interface of Bio/nano, Department of Chemistry and the McKnight Brain Institute, University of Florida, Gainesville, FL 32611, USA.
Tan
Weihong
W
D001483:Base Sequence; D017931:DNA Primers; D015342:DNA Probes; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D012680:Sensitivity and Specificity
D017931:DNA Primers; D015342:DNA Probes