Hybridization of labeled RNA to DNA in agarose gels.
Hybridization of labeled RNA to DNA in agarose gels.
复制标题
标记的 RNA 与 DNA 在琼脂糖凝胶中杂交。
作者：
T. Shinnick;E. Lund;O. Smithies;F. Blattner
在琼脂糖凝胶中电泳后，可以通过在凝胶中（原位）与放射性同源 RNA 杂交来鉴定特定的 DNA 限制性内切酶片段。 RNA-DNA 杂交体通过凝胶放射自显影来检测。比较放射自显影图和溴化乙锭染色凝胶的条带图案可以鉴定与 RNA 探针互补的 DNA 片段。该技术快速、简单且廉价。它足够灵敏，可以检测复杂细胞 DNA 的限制性内切酶消化产生的片段混合物中的单个基因。我们使用这种技术来确定 phi80d3ilv+su+7 和大肠杆菌 DNA 的 Hin III 和 Eco R1 片段中哪些包含大肠杆菌的 5S、16S 和 23S 核糖体 RNA (rRNA) 基因。
Specific DNA restriction endonuclease fragments can be identified after electrophoresis in agarose gels by hybridization in the gel (in situ) to radioactive homologous RNA. RNA-DNA hybrids are detected by autoradiography of the gel. Comparison of band patterns of the autoradiogram and the ethidium bromide stained gel allows the identification of the DNA fragment which is complementary to the RNA probe. The technique is rapid, easy and inexpensive. It is sensitive enough to detect individual genes in a mixture of fragments produced by restriction enzyme digestion of complex cellular DNA. We have used this technique to determine which of the Hin III and Eco R1 fragments of phi80d3ilv+su+7 and E. coli DNAs contain the 5S, 16S and 23S ribosomal RNA (rRNA) genes of E. coli.