Step Seven
After polymerization, carefully remove the comb and rinse the wells with water to remove any unpolymerized acrylamide. Clamp the gel onto the gel apparatus and place a sample well-template over the gel to identify the bottom of the sample wells. Fill the lower reservoir with 1 x electrode buffer so that it covers the bottom of the gel. Fill the upper reservoir with electrode buffer so that the wells are filled with buffer.
The gel should be pre-eclectrophoresed if the gel is
to be used for protein purification prior to sequencing. This is accomplished by including 1 mM sodium thioglycolate in the upper buffer during this step. The thioglycolate acts as a scavenger and reacts with unpolymerized acrylamide as it moves with the ion front thereby removing harmful acrylamide from the path of the migrating proteins. If the gel will be used for simple analysis this step may be omitted.
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