Bio-Rad Sub-Cell® Model 192 Cell - Manual (Page 10)

Table 2.2 Gel Volume Requirements
Gel Size 0.5 cm thick 0.75 cm thick 1.0 cm thick
Bases
25 x 10 cm (Model 96) 125 ml 185 ml 250 ml
25 x 15 cm (Model 192) 185 ml 280 ml 375 ml
Trays
25 x 10 cm 125 ml 185 ml 250 ml
25 x 15 cm 185 ml 280 ml 375 ml
25 x 20 cm 250 ml 375 ml 500 ml
25 x 25 cm 310 ml 465 ml 625 ml
2. Add the agarose to a suitable container (e.g., 500-ml Erlenmeyer flask, Wheaton bottle, etc.).
Add the appropriate amount of electrophoresis buffer (see Section 3) and swirl to sus-
pend the agarose powder in the buffer. If using an Erlenmeyer flask, invert a 50 ml
Erlenmeyer flask into the open end of the 500-ml Erlenmeyer flask containing the agarose.
The small flask acts as a reflux chamber, thus allowing long or vigorous boiling without
much evaporation.
Note: Place a mark on the flask at the liquid level. If evaporation occurs, water can be
added to bring the volume back to the original liquid level.
3. The agarose can be melted by boiling on a magnetic hot plate or in a microwave oven.
Caution: Always wear protective gloves, goggles, and a lab coat while preparing and
casting agarose gels. Boiling molten agarose or the vessels containing hot agarose can
cause severe burns if allowed to contact skin. Molten agarose can become super-heated
and boil over vessels when swirled which can also cause severe burns.
Magnetic Hot Plate Method
4a. Add a stir bar to the undissolved agarose solution. Heat the solution to boiling while
stirring on a magnetic hot plate. Use the appropriate size container to allow bubbles or
foam to disrupt before rising to the neck of the container.
5a. Boil the solution until all of the small translucent agarose particles are dissolved. Set aside
to cool to 50-60 °C before pouring.
Microwave Oven Method
4b. Place the gel solution into the microwave. Using a low to medium setting, set the timer for
a minimum of 5 minutes, stopping the microwave oven every 30 seconds and swirling the
container gently to suspend the undissolved agarose. This technique is the fastest and
safest way to dissolve agarose.
5b. Boil and swirl the solution until all of the small translucent agarose particles are dissolved.
Set aside to cool to 50-60 °C before pouring.
2.2 Comb Set-up
Comb and Comb Holder Set-up
The comb holder used for the Model 96 and 192 was designed to incorporate all the
necessary features required for any agarose gel application. The comb holder allows for
adjustable comb height and can be adjusted so that the comb can be placed anywhere on the
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