title: Development of a simple and effective silver staining protocol for detection of DNA fragments.
abstract: Silver staining is one of the widely used methods for DNA fragment detection in biological research. Silver staining protocols have been steadily optimized to improve detection efficiency. This research reports a continuous effort to simplify the existing silver staining protocols, lower experiment cost, and improve DNA detection sensitivity and image clarity. The new method only requires three reagents (silver nitrate, sodium hydroxide, and formaldehyde) and 6-7 min with high detection sensitivity to visualize as low as 14.6 pg (3.3 pg/mm2) of DNA in a non-denaturing polyacrylamide gel. In comparison to previous reported protocols, the new one has the highest resolution, is the easiest to operate, takes the shortest time, and uses the fewest chemical reagents. Therefore, the new method can be used for quick generation of high quality molecular marker data in genetic analysis.
authors.affiliation: College of Life Sciences, Guangzhou University, Guangzhou, P. R. China.
authors.forename: Wenjie
authors.initials: W
authors.lastname: Liu
authors.affiliation: College of Life Sciences, Guangzhou University, Guangzhou, P. R. China.
authors.forename: Ronghua
authors.initials: R
authors.lastname: Li
authors.affiliation: College of Life Sciences, Guangzhou University, Guangzhou, P. R. China.|Debre Markos University, College of Agriculture and Natural Resources, Debre Markos, Ethiopia.
authors.forename: Habtamu
authors.initials: H
authors.lastname: Ayalew
authors.affiliation: College of Life Sciences, Guangzhou University, Guangzhou, P. R. China.
authors.forename: Yanshi
authors.initials: Y
authors.lastname: Xia
authors.affiliation: Hard Winter Wheat Genetics Research Unit, United States Department of Agriculture - Agricultural Research Service, Manhattan, KS, USA.
authors.forename: Guihua
authors.initials: G
authors.lastname: Bai
authors.affiliation: The USA Institute of Agriculture, The University of Western Australia, WA, Australia.|School of Plant Biology, Faculty of Science, The University of Western Australia, Perth, WA, Australia.
authors.forename: Guijun
authors.initials: G
authors.lastname: Yan
authors.affiliation: The USA Institute of Agriculture, The University of Western Australia, WA, Australia.
authors.forename: Kadambot H M
authors.initials: KHM
authors.lastname: Siddique
authors.affiliation: College of Life Sciences, Guangzhou University, Guangzhou, P. R. China.
authors.forename: Peiguo
authors.initials: P
authors.lastname: Guo
mesh_terms: D000180:Acrylic Resins; D004247:DNA / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D007202:Indicators and Reagents; D012680:Sensitivity and Specificity; D016622:Silver Staining / Q000379:methods*
chemical_list: D000180:Acrylic Resins; D007202:Indicators and Reagents; C016680:polyacrylamide gels; D004247:DNA
keywords: DNA; Non-denaturing polyacrylamide gel; Silver staining