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SDS PAGE: problem with acrylamine and bisacrylamide - (Mar/17/2008 )

excl.gif I have just started working with SDS PAGE. but when i dissolve bisacrylamide (1%) after dissolving acrylamide (29%) the solution becomes turbid and gel does not solidify. All the other chemicals TMED, APS , and other buffere solutions are working properly. Bisacryamide and acrylamide both are not expired. Can anyone tell why it is so and how to overcome this problem?

-fabi-

Just stir it for long time.

-genehunter-1-

set the solution in a pan of warm water while stirring, it will dissolve faster.

-smu2-

When preparing acryl-bis solution let it stir for more than 30 min, always use an ambar bottle for storage and use it within a month. If a gel don't polimeryze could be: not enough TEMED and ammonium persulfate, reagents too old. The APS should be prepare as fresh as posible. I usually prepare small quantity and use within a week (store at 4C). Let the gel polymerize at least 1 hour. If preparing a discontinue gel when adding the buthanol use just a thin layer and wash it with ddH2O (alcohol will inhibit the polymerization of the stacking gel).

-merlav-

QUOTE (merlav @ Mar 18 2008, 12:25 AM)
When preparing acryl-bis solution let it stir for more than 30 min, always use an ambar bottle for storage and use it within a month. If a gel don't polimeryze could be: not enough TEMED and ammonium persulfate, reagents too old. The APS should be prepare as fresh as posible. I usually prepare small quantity and use within a week (store at 4C). Let the gel polymerize at least 1 hour. If preparing a discontinue gel when adding the buthanol use just a thin layer and wash it with ddH2O (alcohol will inhibit the polymerization of the stacking gel).

Thanks alot.

-fabi-