Chemiluminescent nucleic acid detection with digoxigenin-labeled probes: a model system with probes for angiotensin converting enzyme which detect less than one attomole of target DNA.
Digoxigenin-labeled nucleic acid probes for angiotensin-converting enzyme were prepared by the polymerase chain reaction. In a model system, these probes were used to optimize conditions for target detection by direct chemiluminescence after dot-blot, Southern, or Northern hybridizations. Digoxigenin-labeled probes detected 23 fg (1.6 X 10(-19) mol or 9 X 10(4) copies) of target consistently.
Lanzillo
J J
JJ
New England Medical Center, Pulmonary Division, Boston, Massachusetts 02111.
D000818:Animals; D001483:Base Sequence; D001710:Biotin / Q000031:analogs & derivatives / Q000378:metabolism; D015152:Blotting, Northern; D015139:Blotting, Southern; D002417:Cattle; D004247:DNA / Q000032:analysis*; D015342:DNA Probes / Q000138:chemical synthesis*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D003856:Deoxyuracil Nucleotides / Q000378:metabolism; D004076:Digoxigenin / Q000031:analogs & derivatives / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D008163:Luminescent Measurements; D008832:Microchemistry / Q000379:methods; D008969:Molecular Sequence Data; D009696:Nucleic Acids / Q000032:analysis*; D007703:Peptidyl-Dipeptidase A / Q000032:analysis*; D016133:Polymerase Chain Reaction / Q000379:methods
D015342:DNA Probes; D003856:Deoxyuracil Nucleotides; D009696:Nucleic Acids; C070202:digoxigenin-11-deoxyuridine triphosphate; D001710:Biotin; C045931:biotin-11-dUTP; D004247:DNA; D004259:DNA-Directed DNA Polymerase; D007703:Peptidyl-Dipeptidase A; D004076:Digoxigenin