A Fast Silver Staining Protocol Enabling Simple and Efficient Detection of SSR Markers using a Non-denaturing Polyacrylamide Gel

被引:18
|
作者
Huang, Ling [1 ]
Deng, Xiaohui [1 ]
Li, Ronghua [1 ]
Xia, Yanshi [1 ]
Bai, Guihua [2 ]
Siddique, Kadambot H. M. [3 ]
Guo, Peiguo [1 ]
机构
[1] Guangzhou Univ, Coll Life Sci, Guangzhou, Guangdong, Peoples R China
[2] USDA ARS, Hard Winter Wheat Genet Res Unit, Washington, DC 20250 USA
[3] Univ Western Australia, UWA Inst Agr, Nedlands, WA, Australia
来源
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | 2018年 / 134期
关键词
Genetics; Issue; 134; Silver staining; polyacrylamide gel; simple and efficient protocol for PCR detection; SSR marker; flowering Chinese cabbage; tobacco; MICROSATELLITE MARKERS; DNA; PLANTS;
D O I
10.3791/57192
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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 the detection of SSR markers in a polyacrylamide gel. However, conventional protocols for silver staining are technically demanding and 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 reduces reagent costs and enhances the detection resolution and picture clarity. The new method requires two major steps (impregnation and development) and three reagents (silver nitrate, sodium hydroxide, and formaldehyde), and only 7 min of processing for a non-denaturing polyacrylamide gel. Compared to previously reported protocols, this new method is easier, quicker and uses fewer chemical reagents for SSR detection. Therefore, this simple, low-cost, and effective silver staining protocol will benefit genetic mapping and marker-assisted breeding by a quick generation of SSR marker data.
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页数:7
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