Elbrecht A, Lazier C B
J Biochem Biophys Methods. 1984 Jul;9(3):215-20. doi: 10.1016/0165-022x(84)90026-5.
We describe a method for hybridization of cDNA probes to RNA directly in agarose gels which provides a practical alternative to methods involving transfer of the RNA out of the gel. Total cellular RNA is subjected to electrophoresis in agarose gels containing methylmercuric hydroxide as the denaturing agent. After removal of the methylmercuric hydroxide, the gel is dried and 32P-labeled DNA probes are hybridized to the immobilized RNA. This method is more economical in time and expense than methods involving transfer of the RNA out of the gel, while maintaining a level of sensitivity comparable to other procedures.
我们描述了一种在琼脂糖凝胶中直接将cDNA探针与RNA杂交的方法,该方法为涉及将RNA从凝胶中转移出来的方法提供了一种实用的替代方法。将总细胞RNA在含有氢氧化甲基汞作为变性剂的琼脂糖凝胶中进行电泳。去除氢氧化甲基汞后,将凝胶干燥,并将32P标记的DNA探针与固定化的RNA杂交。与涉及将RNA从凝胶中转移出来的方法相比,该方法在时间和费用上更经济,同时保持与其他方法相当的灵敏度水平。