Bio-Rad DCode™ Universal Mutation Detection System - Manual (Page 75)

TTGE Gels
1. The electrophoresis tank should contain 7 L of 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 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.
6. When the set initial temperature is reached, press the °C/RR button to select the ramp rate. Set
the desired ramp rate using the raise and lower buttons. Allow the heater to equilibrate to the
lower ramp rate for 5–10 minutes before loading samples. Press the °C/RR button to return to
the temperature readout.
Heteroduplex, CSGE, and PTT Gels
1. The electrophoresis tank should contain at least 2 L of the appropriate running buffer.
2. 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.
8.2 Sample Loading
1. Remove the clear loading lid. Wash the wells with running buffer to remove any unpoly-
merized gel material or denaturants in the wells.
2. Load the samples using a pipetman and a sequencing loading tip. Be careful not to pierce
the wells during sample delivery.
3. Place the clear loading lid on top of the temperature control module.
8.3 Running the Gel
1. Attach the electrical leads to a suitable DC power supply. Recommended power supply:
Bio-Rad’s Power Pac 300 or 3000.
DGGE, CDGE, and TTGE Gels
1. For DGGE and CDGE run the gel at 130 volts. Apply power to the DCode system and
begin electrophoresis. As a precaution, always set the voltage, current, and power limits
when possible.
Note: The voltage should not exceed 180 V, electrophoretic heating may affect results.
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