ONE-HOUR DOWNWARD ALKALINE CAPILLARY TRANSFER FOR BLOTTING OF DNA AND RNA
ONE-HOUR DOWNWARD ALKALINE CAPILLARY TRANSFER FOR BLOTTING OF DNA AND RNA
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DOI：
10.1016/0003-2697(92)90185-a
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发表时间：
1992-02-14
影响因子：
2.9
通讯作者：
CHOMCZYNSKI, P
中科院分区：
文献类型：
--
作者：
CHOMCZYNSKI, P
使用含有3 mNaCl和8 mNaOH的转移溶液进行DNA和RNA从琼脂糖凝胶到杂交膜的向下碱性毛细管转移。在温和的碱性条件下，DNA和RNA从琼脂糖凝胶上完全洗脱，并在1小时内结合到杂交膜上。在此基础上，提出了一种新的转移方法--向下碱性印迹法。它为常用的Southern和北方印迹方案提供了一种快速有效的替代方案。DNA和RNA的向下碱性印迹可分别在2.5和1.5小时内完成，并且可与塑料和硝酸纤维素膜一起使用。此外，向下碱性印迹方案允许DNA和RNA的杂交效率高于在中性pH下进行的标准印迹方案。
The downward alkaline capillary transfer of DNA and RNA from agarose gel to a hybridization membrane was performed using a transfer solution containing 3 m NaCl and 8 mm NaOH. Under mild alkaline conditions, DNA and RNA were completely eluted from the agarose gel and bound to a hybridization membrane within 1 h. On the basis of this new method of transfer a blotting protocol, downward alkaline blotting, was elaborated. It provides a fast and efficient alternative to commonly used Southern and Northern blotting protocols. The downward alkaline blotting of DNA and RNA can be completed in 2.5 and 1.5 h, respectively, and can be used with both plastic and nitrocellulose membranes. In addition, the downward alkaline blotting protocol allows for a hybridization efficiency of DNA and RNA higher than that of the standard blotting protocols performed at neutral pH.