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30% Acrylamide (100 ml)

  1. Weigh out 29 g acrylamide and 1 g N,N?/EM>-methylenebisacrylamide.
    <CAUTION> Acrylamide is a potent neurotoxin and is absorbed through the skin. The effects of acrylamide are cumulative. Wear glovesand a mask when weighing powdered acrylamide and methylenebisacrylamide. Wear gloves when handling solutions containing these chemicals. Although polyacrylamide is considered to be non-toxic, it should be handled with carebecause of the possibility that it might contain small quantities of unpolymerized acrylamide.
  2. Add 60 ml distilled water.
  3. Heat the solution to 37 C to dissolve the chemicals.
  4. Make upvolume to 100 ml with distilled water.
    <CRITICAL STEP> Cheaper grades of acrylamide and bisacrylamide are often contaminated with metal ions. Stock solutions of acrylamide can easily be purified by stirring overnight with about 0.2 volume of monobed resin (MB-1, Mallinckrodt), followed by filtration through Whatman No. 1 paper.
  5. Sterilize the solution by filtration through a Nalgene filter (0.45- micron pore size).
  6. Check that the pH of the acrylamide solution is 7.0 or less.
    Store the solution in dark bottles at 4 C. During storage, acrylamide and bisacrylamide are slowly converted to acrylic acidand bisacrylic acid, so solutions that are older than 1 year should be discarded

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