Electrophoresis
Development & Consulting
Dr. Hanspeter Schickle
Wolfgang Gstrein

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We use this staining protocol because it will fix also the smaller DNA-fragments below 150 bp!
Please see what happens if weaker fixing recipes are used: “Silver Trouble”
This staining procedure give also best results with denaturing, 7 M Urea containing gels!

Our Stock Solutions: (order numbers of functioning chemicals see page: General Silver Staining)

Fixing Concentrate (5x): 15 g Benzene sulfonic acid (= Phenyl sulfonic Acid) + 120 ml ethanol.
Fill up to 500 ml with dist. water.
Fixing diluter: 150 ml ethanol. Fill up to 500 ml with dist. water.
Washing Concentrate (6x): 1.75 g Benzene sulfonic acid. Fill up to 500 ml with dist.water.
Silvering Concentrate (5x): 5 g AgNO₃ + 1.75 g Benzene sulfonic acid.
 Fill up to 500 ml with dist water.
Developing Concentrate (5x): 62.5 g Na₂CO₃. Fill up to 500 ml with dist. water.
Thiosulfate Concentrate: 500 mg Sodiumthiosulfate (* 5 H₂0). Fill up to 25 ml with dist. water. Renew this solution once per week.

The Final Solutions:

Fixing solution: 40 ml Fixing Concentrate + 160 ml Fixing Diluter
Washing solution: 100 ml Washing Concentrate +500 ml water
Silvering solution: 40 ml Silvering Concentrate + 160 ml dist. water + 260 ul Formaldehyde.
Developing solution: 40 ml Developing Concentrate + 160 ml dist. water + 260 ul Formaldehyde+ 200 ul Thiosulfate Concentrate
 Stopping and preserving: 75 ml acetic acid + 75 ml glycerol + 600 ml dist.water

EDC Electrophoresis Development & Consulting, Vor dem Kreuzberg 17, 72070 Tübingen (Germany)