A Method for Sensitive Staining of DNA in Polyacrylamide Gels Using Basic Fuchsin

Abstract

Background: PAGE is a widely used analytical method to resolve components of a DNA mixture based on their size. Various DNA visualization methods including fluorescence, visible dye and silver have been used for the detection of gel-separated DNA, with each having different advantages and disadvantages in terms of sensitivity, safety and simplicity. Results: A fast and sensitive visible dye-based staining method for DNA in polyacrylamide gels using basic fuchsin (BF) is described. As low as 10–20 pg of DNA can be visualized within 10 min; the sensitivity is fourfold more sensitive than that of SYBR^® Gold stain, the most sensitive commercial fluorescent probe, but similar to silver staining kit from GE Healthcare. In addition, the mechanism studies suggest that the interaction of BF with DNA is mainly contributed by non-intercalative binding mode. Conclusion: By comprehensive studies of this visible dye-based protocol, we concluded that BF stain is a fast and sensitive method currently available for detecting DNA in polyacrylamide gels.

Financial & competing interests disclosure

This work was supported by the National Key Scientific Instrument Development Program of China (2011YQ030139); the Development Program of China (863 Program) (2011AA02A114); and the project sponsored by the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry, Zhejiang Provincial Program for the Cultivation of High-Level Innovative Health Talents and the Science and Technology Program of Wenzhou (Y20090242, S20090031 and Y20110029). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.

Additional information

Funding

This work was supported by the National Key Scientific Instrument Development Program of China (2011YQ030139); the Development Program of China (863 Program) (2011AA02A114); and the project sponsored by the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry, Zhejiang Provincial Program for the Cultivation of High-Level Innovative Health Talents and the Science and Technology Program of Wenzhou (Y20090242, S20090031 and Y20110029). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.