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problem
possible cause
Gel sandwich
Dirty or damaged
leaks while
components
casting
Mis-aligned parts
Over-clamping
Sample
Air bubbles
wells
damaged or
irregular
Incomplete or delayed
polymerization
Debris in wells
Comb removal
Incomplete
Chemicals
gel polymer-
ization
•
p26
Troubleshooting
remedy
Plates, spacers, and the gasket must be completely
clean. Wash if necessary.
Replace chipped plates (especially if chipped near the
spacers).
Check the caster gasket for cuts or cracks and replace if
necessary.
Check plate and spacer alignment, realign if necessary.
Turn cam only as far as necessary to create a seal (usu-
ally 90–150°, but up to 180°).
On each spacer apply a light film of Gel Seal compound
to the bottom outside corner only. Do not use silicone
grease.
Remove air bubbles before inserting combs. Slide comb
into solution at an angle. If comb must be removed, add
more monomer solution before reinserting the comb.
Allow acrylamide gels to set for a minimum of 1 h.
Rinse out unpolymerized gel with sample buffer.
Remove the comb at a slight angle and very slowly to pre-
vent damaging the gel.
Agarose gels: Lower the comb no more than 1 cm into
the gel.
Use only recent stocks of the highest-quality reagents.
If the dry ammonium persulfate does not crackle when
added to water, replace with fresh stock
Increase TEMED or APS concentration, or both.
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