Cut off the top corner of the high numbered end of gel for orientation. We sometimes use a multiple gel holder from Promega for processing several gels at a time. Two gels require the same volume of solution as one. Rinse the gel 3 times for 2 minutes each with distilled water with gentle agitation. Add staining solution to gel and gently agitate for 30 minutes, decant and precipitate silver.
Longer rinses result in weaker signal.
If the rinse goes too long, repeat step 5 with the staining solution. Transfer the gel to 800 mls of chilled developing solution and gently agitate with a rocking motion until the bands first become visible, usually within in a few minutes. Decant the used developer, add the remaining chilled developer and continue to gently agitate until the bands have reached desired intensity. Over-development will result in a brown background with low contrast bands rather than a pencil gray background with sharp bands. Lift up plate to ensure that the neutralizing solution comes in contact with developer underneath the plate.