Gels thinner than 0.4mm may give weak signals while gels thicker than 0.4mm or higher percentage gels may require longer processing times. Also, thicker and high percentage gels are more likely to crack during drying. Do not use wedge gels- they stain unevenly. In contrast to autoradiography, silver stained gels are processed while one surface remains attached to a gel plate. To facilitate this one plate must be silicone treated with an agent such as SigmaCote for the gel to release while the other plate must be silanized with Bind Silane for the gel to adhere.
Silver staining is carried out with the sequencing gel attached to the long glass plate.
Treatment of the long plate with SigmaCote prior to each use is not necessary while the short glass plate should be treated with Bind Silane before each use. GelBond film interferes with the staining of the gels and is not recommended. We have identified several parameters which are critical for successful silver staining. Use only the sodium carbonate provided or ACS reagent grade. The length of the rinse step following staining is critical.