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silver staining

Douglas Rhoads DRHOADS at MERCURY.UARK.EDU
Mon Jul 3 08:58:08 EST 1995


> To:            diagnost at net.bio.net
> From:          lpereira at usp.br (Lygia da Veiga Pereira)
> Subject:       silver staining
> Date:          27 Jun 1995 10:32:58 -0700

> 
> I have tryed a few methods for silver staining DNA in polyacrilamide gels, 
> but they all give me high background.  Can anyone send me their protocol for 
> this technique?
> 
> Thanks!
> 
> Lygia Pereira, Ph.D.
> Universidade de Sao Paulo, Brazil


Silver Stain for Nucleic Acids
     from M. Balas, Volcani Centre, Rehovot, Israel via E. Rybicki Univ. of Cape Town
Short Procedure
1)   Soak in 30% EtOH 10% HOAc 10' room temp. no shaking (can go overnight)
2)   Soak 2x 10' in 10% EtOH
3)   Wash 3x 5' in H2O
4)   Soak 30' in 6 mM AgNO3 (50 ml/gel??) the quick rinse with H2O
5)   Rinse rapidly in fresh 2.5% Na2CO3 0.02% HCHO then replace with fresh for 2' more. 
     Replace with fresh solution and allow bands to develop.
6)   Stop with 1% HOAc and store in same.  Can destain with 2% NH4peroxydisulphate. 
     Gels can be shrunk with 100% EtOH, then acetone and then air dried.
Long Procedure
1)   Soak gel in 50% EtOH 10% HOAc 1-3 hrs or overnight
2)   Soak in 10% EtOH 1% HOAc 1 hr. then rinse 3x with H2O
3)   Soak in 12 mM AgNO3 1 hr. rinse 3x 5' in H2O
4)   Rinse rapidly in fresh 0.75 M KOH 0.28% HCHO, then replace with fresh and allow
     bands to develop
5)   Stop with 0.07 M Na2CO3 or 5% HOAc.  Store in 1% HOAc.
 
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||Doug Rhoads              || Dept. of Biological Sciences||
||drhoads at mercury.uark.edu || 601 Science Engineering     ||
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