| | |||||||
| Register | Search | Today's Posts | Mark Forums Read |
| Protocols and Methods Forum Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique. |
| | LinkBack | Thread Tools | Display Modes |
|
#1
| |||
| |||
| I use Silver Stain for RNa and DNA in Polyacrylamide Gels and in Agarose gel. In the Polyacrylamide gels i use the "Merril" methods (Science, vol. 211,1427-1428, 27 march 1981) that was write for Proteins(in bief: fix the gel, rins the gel, soak in a solution of potassium dichromate, wash the gel, soak in a solution of silver nitrate, rins in developer solution until the desired inensity, stop solution) but work very well also for nucleic acids. Instead to fix the gel in the 50 % methanol and 12% acetic acid I use 40% ethanol- 10% acetic acid for 20'. After i use 9% ethanol-1% acetic acid to conserve the gel at 4°C before Stains, but realy I don't know wich are the best solution for fix and conserve the gel before Stain (Someone can help me?) . Besides I use Ethanol instead of methanol only becouse is more cheap and less toxic, but i don't know why Merril instead suggest methanol (Why methanol?). Often the stain is more yellow than black, why? Do it hit with ethanol? In the agarose gel i try to use the methods of Zalazar et all*(to see the note) with little result for the background. The methods is this: submerge the gel in 18 mM Silver nitrate- 10% acetic acid for 10' or more; to rins in wather more times; to develop in 3% NaOH-o,5 mL/L of 37% formaldehyde- 2mg/L sodium thisulfate; stop solution (10%acetic acid). Someone use this method or other methods for obtain a good Silver Stain of DNA in Agarose gels (the problem is the background)? I've read too that is possible to drying the gels with cellophane sheets, but it's not clear if i can do this (withaut new machine ), how i can do this and if drying the gel can be useful to improve the stain. Sorry for my english, i hope to be clear, thanks . -- Fabio FabioDB#deleteme#@katamail.com ----------------------------------------------- Note: *Modified Silver Staining for RNA and DNA in Agarose Gels. Analytical Biochemistry 291, 299-300 (2001) and A Procedure of Silver Staining for Nucleic Acids in Agarose Gels without Pretreatment or Drying Steps, Analytical Biochemistry 252, 15-18 (1997) |
| Tags |
| questions , silver , stain |
| Thread Tools | |
| Display Modes | |
|
|
| | ||||
| Thread | Thread Starter | Forum | Replies | Last Post |
| nonspecific bands in silver stain IPs | nshaikh | Immunoprecipitation Forum | 0 | 09-30-2009 05:33 PM |
| good morning | Tim Fitzwater | Protocols and Methods Forum | 0 | 04-16-2008 02:21 PM |
| DNA Visualization Using Silver Stain | galgur | DNA Techniques | 1 | 02-20-2008 11:32 AM |
| ladder for silver stain | Marzia Barbieri | Protocols and Methods Forum | 0 | 08-31-2005 10:15 AM |
| Sci.chem FAQ - Part 7 of 7 | Bruce Hamilton | Chemistry Forum | 0 | 01-15-2004 08:13 AM |