Skip to main content
ARS Home » Plains Area » Manhattan, Kansas » Center for Grain and Animal Health Research » Hard Winter Wheat Genetics Research » Research » Publications at this Location » Publication #348269
Title: A fast silver staining protocol enabling simple and efficient detection of SSR markers in a non-denaturing polyacrylamide gel

Author
item HUANG, LIN - Guangzhou University
item DENG, XIAOHUI - Guangzhou University
item LI, RONGHUA - Guangzhou University
item XIA, YANSHI - University Of Western Australia
item Bai, Guihua
item SIDDIQUE, KADAMBOT H.M. - University Of Western Australia
item GUO, PEIGUO - Guangzhou University

Submitted to: Journal of Visualized Experiments
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/1/2018
Publication Date: 4/20/2018
Citation: Huang, L., Deng, X., Li, R., Xia, Y., Bai, G., Siddique, K., Guo, P. 2018. A fast silver staining protocol enabling simple and efficient detection of SSR markers in a non-denaturing polyacrylamide gel. Journal of Visualized Experiments. 134:E57192(1-7). https://doi.org/10.3791/57192.
DOI: https://doi.org/10.3791/57192

Interpretive Summary: Silver staining of polyacrylamide gels is a commonly used method for DNA marker visualization, especially for simple sequence repeat (SSR) markers. Currently available protocols for silver staining are complicated and time-consuming. In this study, we simplified the silver staining method by reducing processing steps and reagents without sacrificing DNA detection resolution. The new method only requires two simple steps and three reagents, which significantly reduced cost and time of DNA staining. A video was made to demonstrate how to use this new method step-by-step. This simple, low-cost and effective silver staining protocol will improve genotyping efficiency in marker-assisted breeding.

Technical Abstract: Simple Sequence Repeat (SSR) is one of the most effective markers used in plant and animal genetic research and molecular breeding programs. Silver staining is a widely used method for detection of SSR markers in a polyacrylamide gel. However, conventional protocols of silver staining involve a high technical demand and are time-consuming. Like many other biological laboratory techniques, silver staining protocols have been steadily optimized to improve detection efficiency. Here we report a simplified silver staining method that significantly lowers reagent cost and enhances detection resolution and picture clarity. The new method only requires two major steps (impregnation and development) and three reagents (silver nitrate, sodium hydroxide and formaldehyde), and uses 7 min processing time for a non-denaturing polyacrylamide gel. Compared with previously reported protocols, this new method is easier and quicker, and uses fewer chemical reagents for SSR detection. Therefore, this simple, low-cost and effective silver staining protocol will benefit gene mapping and marker-assisted breeding by quick generation of SSR marker data.