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一种用于聚丙烯酰胺凝胶中核酸染色的简化二氨银法。

A simplified silver diammine method for the staining of nucleic acids in polyacrylamide gels.

作者信息

Vari F, Bell K

机构信息

Queensland Institute of Medical Research, Royal Brisbane Hospital, Australia.

出版信息

Electrophoresis. 1996 Jan;17(1):20-5. doi: 10.1002/elps.1150170104.

Abstract

High sensitivity and low background, which are the significant features of the procedure of Johansson and Skoog (J. Biochem. Biophys. Methods. 1987, 14, (Suppl.) 33) for silver staining of nucleic acids in polyacrylamide gels, have been improved by excluding ethanol from all solutions and increasing the length of washing steps after sensitisation and staining. Including a fixation step before sensitisation increased the contrast of the bands even at high concentration of nucleic acid, producing a shift in the tone of colour from black to grey instead of gold to yellow bands. This technique is superior in terms of sensitivity and background to a number of other silver staining protocols, three which utilise silver diammine and three which utilise silver nitrate ions.

摘要

高灵敏度和低背景是约翰松和斯科格(《生物化学与生物物理方法杂志》,1987年,第14卷,(增刊)33)在聚丙烯酰胺凝胶中对核酸进行银染的方法的显著特征,通过从所有溶液中排除乙醇并增加敏化和染色后洗涤步骤的时长,这些特征得到了改进。在敏化前加入固定步骤,即使在核酸浓度较高时也能增加条带的对比度,使颜色色调从黑色变为灰色,而不是从金色变为黄色条带。在灵敏度和背景方面,该技术优于其他一些银染方案,其中三种使用二氨合银,三种使用硝酸银离子。

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