Lund V, Lindqvist B H, Eggset G
Institute of Medical Biology, University of Tromsø, Norway.
Nucleic Acids Res. 1989 Jan 25;17(2):539-51. doi: 10.1093/nar/17.2.539.
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.
这种用于检测核酸的非同位素方法基于通过紫外线照射对核酸进行原位标记。不同的紫外线诱导光产物，主要是胸腺嘧啶二聚体型，可被针对所引入损伤的纯化兔抗体识别。然后，通过常规免疫染色程序可使紫外线标记的核酸可视化。该技术的一个主要优点是成本低且DNA特异性标记简便。已证明纯化的兔抗体对紫外线照射的DNA具有特异性，并且该方法已用于检测与膜支持物上同源靶DNA杂交的特定DNA序列。我们认为该方法的灵敏度可以提高，并讨论了使用不同紫外线剂量、免疫染色方法和膜类型的意义。
