Problems with SDS-PAGE

JOHN PROUDMAN jproudman at ASRR.ARSUSDA.GOV
Thu Apr 21 16:49:18 EST 1994


Paolo, I think that your problem with the poor shape of the sample wells in
your gels is due to incomplete polymerization of the polyacrylamide at the
top of your gel.  This is due to oxygen inhibiting polymerization where the
comb/top of the gel is exposed to air.  Not knowing what kind of apparatus
you are using, it is difficult to recommend an exact fix, but you need to
exclude oxygen (as much as possible) from the gel until it is polymerized.
I do this by placing my casting stand into the empty buffer chamber, putting
the top on, and flushing out the air by a stream of nitrogen.  Good luck.
John




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