Use of stable dye-DNA intercalating complexes to detect cystic fibrosis mutations.
Several dyes are now known which intercalate into double-stranded DNA with fluorescence enhancement. One such is TOTO (1,1'-(4,4,7,7-tetramethyl-4,7-diazaundecamethylene)-bis-4-[3-meth yl-2,3- dihydro-(benzo-1,3-thiazole)-2-methlyidene]-quinolinium tetraiodide), a homodimer of thiazole orange, whose complexes with DNA are stable to electrophoresis and show a 1000-fold enhancement after binding. We demonstrate that this reagent can be successfully used in combination with a laser-excited fluorescence gel scanner to detect the three most common cystic fibrosis mutant alleles, delta F508, G551D and G542X. This has potential application to mass population heterozygote screening.
Human Genetics Unit, University of Edinburgh, Western General Hospital, UK.
R A
RA
Axton
D J
DJ
Brock
D000483:Alleles; D001483:Base Sequence; D003550:Cystic Fibrosis / Q000175:diagnosis* / Q000235:genetics / Q000517:prevention & control; D004247:DNA / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D005453:Fluorescence; D006579:Heterozygote; D006801:Humans; D008403:Mass Screening; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D011485:Protein Binding; D011806:Quinolinium Compounds / Q000378:metabolism*; D013844:Thiazoles / Q000378:metabolism*
D011806:Quinolinium Compounds; D013844:Thiazoles; C075295:1,1'-(4,4,7,7-tetramethyl-4,7-diazaundecamethylene)bis-4-(3-methyl-2,3-dihydro(benzo-1,3-thiazole)-2-methylidene)quinolinium; D004247:DNA