TY - JOUR
T1 - A fast silver staining protocol enabling simple and efficient detection of ssr markers using a non-denaturing polyacrylamide gel
AU - Huang, Ling
AU - Deng, Xiaohui
AU - Li, Ronghua
AU - Xia, Yanshi
AU - Bai, Guihua
AU - Siddique, Kadambot H.M.
AU - Guo, Peiguo
PY - 2018/4/20
Y1 - 2018/4/20
N2 - 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.
AB - 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.
KW - Flowering Chinese cabbage
KW - Genetics
KW - Issue 134
KW - Polyacrylamide gel
KW - Silver staining
KW - Simple and efficient protocol for PCR detection
KW - SSR marker
KW - Tobacco
UR - http://www.scopus.com/inward/record.url?scp=85046628356&partnerID=8YFLogxK
U2 - 10.3791/57192
DO - 10.3791/57192
M3 - Article
C2 - 29733306
AN - SCOPUS:85046628356
VL - 2018
JO - JOURNAL OF VISUALIZED EXPERIMENTS : JOVE
JF - JOURNAL OF VISUALIZED EXPERIMENTS : JOVE
SN - 1940-087X
IS - 134
M1 - e57192
ER -