A rapid and effective method for silver staining of PCR products separated in polyacrylamide gels.
With the development of molecular quantitative genetics, particularly, genetic linkage map construction, quantitative trait loci mapping or genes fine mapping and association analysis etc., more and more PCR products separated in polyacrylamide gels need to be silver-stained. However, conventional silver-staining procedures are complicated and time-consuming as they require a lot of preparation and handling of several solutions prior to use. In this study, a simple and rapid protocol for silver staining of PCR products was developed. The number of steps was reduced compared to conventional protocols, thus achieving detection of PCR products in 7 min, saving time and resources. Fixation and staining solution and developing solution in present staining procedure allowed a reutilization for 12 and 8 times, respectively, reducing the cost greatly. Meanwhile, the sensitivity was significantly improved with the improved method and the minimum of 0.097 ng/μL of DNA amount can be detected in denaturing polyacrylamide gel. The protocol developed in this study will facilitate the development of molecular quantitative genetics.
National Field Genebank for Tropical Fruit (Zhanjiang), South Subtropical Crops Research Institutes, Chinese Academy of Tropical Agricultural Sciences, Zhanjiang, P. R. China.
Qingzhi
Q
Liang
Dingqing
D
Wen
Jianghui
J
Xie
Liqin
L
Liu
Yongzan
Y
Wei
Yicheng
Y
Wang
Shengyou
S
Shi
D001391:Azo Compounds; D004247:DNA / Q000032:analysis* / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D016622:Silver Staining / Q000379:methods*
D001391:Azo Compounds; D004247:DNA; C014527:Eriochrome Black T
Eriochrome black T; PCR products; Polyacrylamide gel; Silver staining