Bio-Rad DCode™ Universal Mutation Detection System - Manual (Page 22)

Pre-heating the Running Buffer
1. Fill the electrophoresis tank with 7 L of 1x TAE running buffer.
Note: It is recommended that the running buffer not be reused. Reusing the running buffer
may affect the migration rate and band resolution.
2. Place the temperature control module on top of the electrophoresis tank. Attach the power
cord to the temperature control module and turn the power, pump, and heater on. The
clear loading lid should be on the temperature control module during preheating.
3. Set the temperature controller to the desired temperature. Set the temperature ramp rate
to 200 °C/hr to allow the buffer to reach the desired temperature the quickest.
4. Preheat the buffer to the set temperature. It can take 1 to 1.5 hours for the system to heat
the buffer up to the set temperature. Heating the buffer in a microwave helps reduce the
preheating time.
Assembling the Perpendicular Gradient Gel Sandwich
For perpendicular gel formats, 7.5 x 10 cm (dual) and 16 x 16 cm (single) gel sandwich sizes
are recommended. These two different perpendicular gel formats consist of a set of spacers that
provide casting at the side of the gel sandwich via the stopcock. To insure proper alignment,
make sure all plates and spacers are clean and dry before assembly. Use caution when
assembling the glass plate sandwiches. Wear gloves and eye protection at all times.
1. Assemble the gel sandwich on a clean surface. Lay the large rectangular plate down first,
then place the left and right spacers of equal thickness along the short edges of the larger
rectangular plate. To assemble perpendicular gradient gels, place the spacers so that the
holes on the spacers are at the top of the plate with the grooved side of the spacer against
the larger glass plate. When properly placed, the notched edges of the spacers will face
each another.
2. Place a short glass plate on top of the spacers so that it is flush with the bottom edge of
the long plate.
3. Loosen the single screw of each sandwich clamp by turning counterclockwise. Place each
clamp by the appropriate side of the gel sandwich with the locating arrows facing up and
toward the glass plates (Figure 4.5).
Fig. 4.5. Positioning glass plates, spacers, and clamps.
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