一种用于聚丙烯酰胺凝胶中核酸的无需固定和预处理的银染程序。

A silver staining procedure for nucleic acids in polyacrylamide gels without fixation and pretreatment.

作者信息

An Z W, Xie L L, Cheng H, Zhou Y, Zhang Q, He X G, Huang H S

机构信息

Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan 571737, China.

出版信息

Anal Biochem. 2009 Aug 1;391(1):77-9. doi: 10.1016/j.ab.2009.04.036. Epub 2009 May 12.

DOI:10.1016/j.ab.2009.04.036

PMID:19442642

Abstract

Silver staining of nucleic acid has been widely used in molecular marker analysis such as simple sequence repeat (SSR), single-strand conformation polymorphism (SSCP), and amplified fragment-length polymorphism (AFLP). Many alternatives to silver staining methods have been described, but these methods are not efficient or cost-effective. Here we report a silver staining method that requires less than 10min for one gel and can save chemicals as well. It has a detection limit of approximately 5.6pg of DNA/mm(2) in nondenaturing polyacrylamide gels and 12.8pg/mm(2) in denaturing polyacrylamide gels.

摘要

核酸银染已广泛应用于分子标记分析，如简单序列重复（SSR）、单链构象多态性（SSCP）和扩增片段长度多态性（AFLP）。已经描述了许多银染方法的替代方法，但这些方法效率不高或成本效益不佳。在此，我们报告一种银染方法，该方法对一块凝胶染色所需时间不到10分钟，并且还能节省化学试剂。在非变性聚丙烯酰胺凝胶中，其检测限约为5.6pg DNA/mm²，在变性聚丙烯酰胺凝胶中为12.8pg/mm²。