Bio-Rad DCode™ Universal Mutation Detection System - Manual (Page 74)

Fig. 8.1. Attaching the sandwich assembly on to the core.
5. Turn the core to its other side and repeat steps 1–4 to attach the second gel sandwich.
Note: When the gel sandwich has been properly installed, the shorter inside glass plate
will be forced against the notch in the U-shaped gasket to create a leak-proof seal. Always
inspect the contact between the gasket and glass plate to make sure the glass plate is
seated against the notch in the gasket and is not resting above or below this notch.
Improper installation of the gel sandwich can result in buffer leakage during the run.
6. If only one gel is to be run, assemble a set of glass plates without the spacers. Place the
short glass plate on top of the long glass plate. Guide the left and right clamps onto the
sandwich so that the plates fit the appropriate notches in the clamp. Insure that the bottom
of the glass plates are flush. Tighten the screws enough to hold the plates in place. No
further alignment is necessary. Attach it to the other side of the core to form an upper chamber
dam.
7. Pour 350 ml of running buffer into the upper buffer chamber. At this point, check the integrity
of the upper buffer seal. If the buffer appears to be leaking, pour the running buffer into a
beaker, remove the gel sandwich assemblies (Section 8.4), re-lubricate the gasket, and
repeat steps 1–4.
DGGE, CDGE, and SSCP Gels
1. The electrophoresis tank should contain 7 L the appropriate running buffer.
2. When the running buffer has reached the desired temperature, turn the system off.
Disconnect the power cord.
3. Remove and place the temperature control module on the DCode lid stand. Place the
core and the attached gel assemblies into the buffer chamber by positioning the red
button towards the right hand side and the black button along the left hand side of the
system. Place the temperature control module on top of the electrophoresis tank.
Note: The core fits into the tank in one orientation only, allowing the core to lock in place.
4. Connect the power cord and turn the power, pump, and heater on. Remove the clear loading
lid and wash the wells with running buffer to remove any unpolymerized gel material/leached
denaturants from the wells. If necessary, add more buffer to the “max” line on the
electrophoresis tank. Place the clear loading lid back onto the temperature control module.
5. Allow the system to reach the set initial temperature before loading samples. This may take
10–15 minutes.
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