Detection of specific DNA sequences using antibodies recognizing UV-labelled DNA.
This non-isotopic method for detection of nucleic acids is based on the in situ labelling of the nucleic acid by exposure to UV-irradiation. The different UV-induced photoproducts, mainly of the thymidine dimer type, are recognized by purified rabbit antibodies specific to the lesions introduced. The UV-labelled nucleic acids can then be visualized by conventional immunostaining procedures. A major advantage of the technique is the low cost and the ease by which the DNA is specifically labelled. The purified rabbit antibodies were shown to be specific for UV-irradiated DNA, and the method was applied for detection of specific DNA sequences hybridized to homologous target DNA on membrane support. We believe that the sensitivity of the method can be improved, and the significance of using different UV-doses, immunostaining methods and membrane types is discussed.
Lund
V
V
Institute of Medical Biology, University of Tromsø, Norway.
Lindqvist
B H
BH
Eggset
G
G
D000818:Animals; D000906:Antibodies; D000937:Antigen-Antibody Reactions; D001482:Base Composition / Q000528:radiation effects; D001483:Base Sequence / Q000528:radiation effects; D015139:Blotting, Southern; D002417:Cattle; D004247:DNA / Q000276:immunology / Q000302:isolation & purification* / Q000528:radiation effects; D015342:DNA Probes; D004307:Dose-Response Relationship, Radiation; D009693:Nucleic Acid Hybridization / Q000528:radiation effects; D011817:Rabbits; D013194:Staining and Labeling; D014466:Ultraviolet Rays
D000906:Antibodies; D015342:DNA Probes; D004247:DNA