Northern blotting of RNA denatured in glyoxal without buffer recirculation
Northern blotting of RNA denatured in glyoxal without buffer recirculation
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DOI：
10.2144/97224st01
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发表时间：
1997-04-01
期刊：
影响因子：
2.7
通讯作者：
Burnett, WV
中科院分区：
文献类型：
--
作者：
Burnett, WV
描述了一种快速简便的制备北方印迹的方法。在溴化乙锭和甘油存在下，用乙二醛使RNA变性，然后在不需要再循环的缓冲液中通过琼脂糖使RNA变性。在没有任何额外洗涤的情况下，将RNA真空印迹到NaOH中的尼龙膜上，同时除去乙二醛加合物。所有这些步骤加上滤膜的预杂交和加入地高辛标记的探针可以在一天内完成。使用标准程序清洗过滤器和检测探针，整个程序可以在两天内完成。
A rapid and easy procedure for preparing Northern blots is described. RNA is denatured with glyoxal in the presence of ethidium bromide and glycerol, then electrophoresed through agarose in a buffer that does not require recirculation. Without any additional washes, the RNA is vacuum-blotted to a nylon membrane in NaOH, which simultaneously removes the glyoxal adducts. All of these steps plus prehybridization of the filter and addition of a digoxygenin-labeled probe can be completed in one day. Using standard procedures to wash the filters and detect the probe, the entire procedure can be completed within two days.