Review the major precautionary steps in preparing a
gel used for protein sequence analysis:
-
Overnight polymerization to reduce unpolymerized
acrylamide.
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Pre-electrophoresis with sodium thioglycolate
acrylamide.
-
Do not boil samples to reduce heat-induced
cleavage.
-
Reduce exposure of gel to acidic and fixing
conditions, ie. stains. This reduces cleavage of internal ASP-PRO peptides bonds and reduced extraction efficiency due to protein fixation.
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