Northern hybridization analysis of RNA using diethylpyrocarbonate-treated nonfat milk.
Northern hybridization analysis of RNA using diethylpyrocarbonate-treated nonfat milk.
复制标题
使用焦碳酸二乙酯处理的脱脂牛奶对 RNA 进行 Northern 杂交分析。
DOI：
10.1016/0003-2697(86)90310-6
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发表时间：
1986
影响因子：
2.9
通讯作者：
Bresnick,E
中科院分区：
文献类型：
--
作者：
Siegel,LI;Bresnick,E
介绍了一种简单且相对廉价的将固定在硝基纤维素上的RNA与放射性DNA探针杂交的方法。该方法是对Johnson等人提出的牛乳胶转移技术优化法的改进。(1984)吉恩·安纳尔。Tech.1，3-8，使用经过核糖核酸酶抑制剂二乙基焦碳酸酯(DEPC)处理的脱脂牛奶，使硝酸纤维素纸上的非特定结合部位饱和，并洗掉未结合的放射性。与标准的Northern杂交条件相比，DEPC-脱脂牛奶杂交技术速度更快、成本更低、更可靠，在放射自显影上产生清晰清晰的RNA条带，几乎没有背景反应性。
A simple and relatively inexpensive method for hybridizing RNA that is immobilized on nitrocellulose to a radioactive DNA probe is presented. The procedure, a modification of the Bovine Lacto Transfer Technique Optimizer method of Johnson et al. (1984) Gene Anal. Tech.1, 3–8, uses nonfat milk which has been treated with the RNase inhibitor, diethylpyrocarbonate (DEPC), to saturate nonspecific binding sites on nitrocellulose paper, and to wash off unbound radioactivity. The DEPC-nonfat milk hybridization technique is faster, less costly, and more reliable than standard Northern blot hybridization conditions, yielding sharp, clear bands of RNA on autoradiographs with virtually no background reactivity.