Thiazole orange as the fluorescent intercalator in a high resolution fid assay for determining DNA binding affinity and sequence selectivity of small molecules.
The viability of using thiazole orange as an alternative to ethidium bromide in a fluorescent intercalator displacement (FID) assay is explored by profiling the DNA binding affinity and sequence selectivity of netropsin. Utilizing a library of hairpin deoxyoligonucleotides containing all possible four base-pair sequences, the method provides a high resolution profile of the DNA binding properties of small molecules in a high throughput format.
Boger
D L
DL
Department of Chemistry and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA. boger@scripps.edu
Tse
W C
WC
D000900:Anti-Bacterial Agents / Q000378:metabolism; D052160:Benzothiazoles; D001667:Binding, Competitive; D004247:DNA / Q000378:metabolism*; D004996:Ethidium / Q000737:chemistry / Q000592:standards; D005456:Fluorescent Dyes / Q000737:chemistry / Q000592:standards*; D015723:Gene Library; D007364:Intercalating Agents / Q000737:chemistry* / Q000592:standards; D008024:Ligands; D008832:Microchemistry; D009429:Netropsin / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000378:metabolism; D011804:Quinolines; D013050:Spectrometry, Fluorescence; D013844:Thiazoles / Q000737:chemistry* / Q000592:standards; D017186:Titrimetry
D000900:Anti-Bacterial Agents; D052160:Benzothiazoles; D005456:Fluorescent Dyes; D007364:Intercalating Agents; D008024:Ligands; D009838:Oligodeoxyribonucleotides; D011804:Quinolines; D013844:Thiazoles; C051000:thiazole orange; D009429:Netropsin; D004247:DNA; D004996:Ethidium