Firestein G S, Gardner S M, Roeder W D
University of California at San Diego School of Medicine 92103.
Anal Biochem. 1987 Dec;167(2):381-6. doi: 10.1016/0003-2697(87)90180-1.
Molecular hybridization with RNA probes was performed on unfractionated cells solubilized in guanidine thiocyanate solutions. Unhybridized probe was digested with ribonuclease, and protected probe fragments were resolved by polyacrylamide gel electrophoresis (PAGE). Since the same medium was used both for solubilization of the cells and as the hybridization buffer, RNA purification was not required and the analysis of large numbers of samples was facilitated. Using this method, specificity is superior to dot blot analysis because the size of hybridized fragments is determined and the signal of the probe hybridized to target RNA is separated from the background by PAGE.
用RNA探针进行分子杂交是在溶解于硫氰酸胍溶液中的未分级细胞上进行的。未杂交的探针用核糖核酸酶消化，受保护的探针片段通过聚丙烯酰胺凝胶电泳（PAGE）分离。由于用于细胞溶解的培养基与杂交缓冲液相同，因此不需要进行RNA纯化，有利于大量样品的分析。使用这种方法，特异性优于斑点印迹分析，因为可以确定杂交片段的大小，并且通过PAGE将与靶RNA杂交的探针信号与背景分离。
