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.
Axton
R A
RA
Human Genetics Unit, University of Edinburgh, Western General Hospital, UK.
Brock
D J
DJ
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