> 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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