Hoefer SE615 - Manual (Page 18)

Pouring stacking gels
You may add stacking gels to the entire set of
polymerized gels —homogeneous or gradient—
while the sandwiches are still in the casting box,
or you may add stacking gel to individual gels
immediately prior to use.
For optimal resolution, add the stacking gel at
the last minute to prevent diffusion of buffers
between the two gel layers.
1
If you are pouring stacking gels later, skip to step 1
under “Removing polymerized gels” on page 17.
If you are pouring stacking gels now, calculate the
volume of stacking gel monomer solution needed by
referring to the following formula:
Monomer vol. = height × width × thickness × total number of
(ml)
(cm) (cm) (cm) stacking gels
Where:
• Height is the distance between the separating gel
and the top of the plates.
• Width is the distance between the spacers.
2
Rinse off the water-saturated n-butanol overlay with
distilled water or Tris buffer and invert the caster to
drain it. Repeat this 2–3 times.
3
Degas the stacking gel monomer solution and add
catalyst and initiator.
•
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