Acrylamide Polymerization A Practical Approach ( contain stacking gels) i.e....)

Acrylamide Polymerization A Practical Approach

contain stacking gels) i.e., any gel which is not overlaid optimal polymerization results when visible gelation takes place in 810 min. Higher initiator concentrations and faster polymerization are required in these cases because of the inhibitory effect of atmospheric oxygen associated with the comb. In any case, conversion of monomer to polymer should be greater than 95%. If gelation takes longer than 20 min, the inhibitory effects of atmospheric oxygen will begin to appear.

As a general rule, use the lowest catalyst concentrations that will allow polymerization in the optimal period of time. In the case of ammonium persulfate/TEMED-catalyzed reactions, for example, approximately equimolar concentrations of both catalysts in the range of 1 to 10 mM are recommended.

Riboflavin is often used as an initiator along with TEMED, or with TEMED and ammonium persulfate. The major advantage of riboflavin is that it is active in very low concentrations (~510 g/ml). Thus, when riboflavin is used with TEMED and ammonium persulfate, the total amount of initiator required (sum of the three initiators) is less. Given the possible effects of initiators on buffer pH, riboflavin-based initiator systems are useful for poorly buffered systems such as electrofocusing gels, in which the only buffering components are ampholytes.

Visible gelation takes longer in riboflavin-based initiator systems, usually 3060 min. Oxygen does not have the dramatic inhibitory effect on riboflavin-based initiator systems that it has on TEMED/ammonium persulfate systems. This is presumably due to the oxygen-scavenging property of riboflavin. As a result, longer gelation time can be tolerated.

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( contain stacking gels) i.e....)Acrylamide Polymerization A Practical Approach