Use of silver staining for DNA in polyacrylamide gels... - or any other helpful suggestios to improve detection! (Feb/17/2008 )

Hi everyone,
Wasn't really sure what forum to post this in, but here we go anyway.....

I am looking into alternatives to Ethidum Bromide for staining my DNA polyacrylamide gels. The safety of EtBr is not an issue, its a matter of sensitivity and detection.

Has anybody used silver staining? I found a kit from Bio-Rad that can be used for both protein and DNA. Has anybody tried this? It claims that it can detect nanogram amounts....I think EtBr is able to detect ~ 10 ng?

Thanks in advance!

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Is it PCR products you want to detect? You could always do a radiolabelled PCR.

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I have been using silver staining quite a lot to visualize DGGE gels (working with DNA). To my knowlenge silver staining provides the best visual result. Nevertheless you might face problems in exhicing bands afterwards. The type of stop solution you will be using as well as the processing of the echiced bands could be critical in order to be able to isolate DNA from the gel, afterwards.

There is quite a lot of literature on silver staining. I could give you some references if you want..

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Thanks for your replies!

Clare: Yes, I am trying to detect PCR products from a very complex sample. I have been looking into end labelling and audioradiography using 35S. Have you used this method? Is it better to label the primer as opposed to end-labelling of the product?

Odiporos: That would be great if you could send me some references. Since I want to analyse PCR products I wouldn't run it on a denaturing gel, probably just on a high-resolution polyacrylamide gel.


I think the autoradiography will be more sensitive but the silver staining would be easier for routine use...

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A quick soft staining method: Mahmood, Freitag and Prosse, 2006, FEMS Microbiol Ecol 56:482–493
An alternative way to retrieve DNA from silver-stained Polyacrylamide Gels: Siembieda and Lakatua, 1998, Clinical Chemistry, 44: 1989-1991

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Nope - haven't used that method. I just did a 'normal' PCR and added in some alpha P32 (dATP) Easy and quick.

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