Help: PAGE sequencing gels

Thu Feb 23 21:51:11 EST 1995

I never had the problem you have. I strongly sure that your troubles
originate not from gel receipe, buffer or smth. other than plates,
spacers, nonuniform thickness of gel because of bad spacers or too
strong clamping. In addition comb thickness should correspond to spacer's.
Somretimes you need to check plates, they are not always uniformly flat,
espacially if you use plates that were produced not in a company like 
BRL, BioRad or Pharmacia. If you see that plates are good, apply 
Sigma Coat or Acrylease (Stratagene) only to one of the plates, other plate
should always be free of that. 
Clamp the plates uniformly and not too strong, otherwise you get the result,
just opposite you awaiting.
Pouring gel the day before is my usual practice. I remove combs just before
experiment. Don't forget to clamp comb area, keeping in mind that it will be
more difficult (more gentle force and patience needed) to remove combs after
1 or 2 days, then if you do it 2 hours after pouring.
I do not remove clamps after polymerization (from any part of the plates)
if I am going to store the gel.
Hope, this will help

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