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吖啶橙荧光增强负染法检测聚丙烯酰胺凝胶中的 DNA

An expanding negative detection method for the visualization of DNA in polyacrylamide gels by eosin Y.

机构信息

Zhejiang Provincial Key Laboratory of Biopharmaceuticals, Wenzhou Medical College, Wenzhou, Zhejiang, PR China.

出版信息

Electrophoresis. 2010 Oct;31(20):3450-6. doi: 10.1002/elps.201000196.

Abstract

A sensitive and easy technique has been developed for the negative detection of DNA following PAGE using eosin Y. After electrophoresis, gels are fixed and stained within 40 min to provide a detection limit of 0.1-0.2 ng of single DNA band, which appears as transparent and colorless under the opaque gel matrix background. The sensitivity of the new stain is fourfold better than zinc-imidazole negative and ethidium bromide stains. Furthermore, the newly developed staining method has been successfully applied to RNA visualization in polyacrylamide gels. In addition, the inclusion of inorganic salts in staining solution was also investigated, which has great effect on the staining efficiency.

摘要

已开发出一种灵敏且易于操作的 PAGE 后检测 DNA 的技术,使用曙红 Y 进行负染。电泳后,凝胶在 40 分钟内固定和染色,可检测到 0.1-0.2ng 单条 DNA 带,在不透明的凝胶基质背景下呈现透明无色。新染色剂的灵敏度比锌咪唑负染和溴化乙锭染色剂高四倍。此外,新开发的染色方法已成功应用于聚丙烯酰胺凝胶中 RNA 的可视化。另外,还研究了染色溶液中无机盐的包含情况,这对染色效率有很大的影响。

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