Instruction manual

Fig. 7.2. Adapting the sandwich assembley 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 butted 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. As a standard procedure, stand

the core and the attached gel sandwiches, pour buffer into the upper buffer chamber, and

check for leaks prior to a run. Pour enough buffer to immerse the cathode wires connected

in the core. If no leaks are present, it is safe to run the gel.

6. If only one gel is to be run, assemble the other 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 appropriate notches in the clamp. 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.

Note: Failure to slide the dam up completely to the top of the clamp will result in buffer

leaking from the upper chamber.

7.2 Loading Samples

1. When the running buffer has reached the correct temperature in the D GENE chamber, the

samples are ready to be loaded onto the gel. Turn the D GENE system off. Disconnect the

power cord. Place the core and the attached gel assemblies into the lower buffer chamber.

The core goes in the chamber in only one direction. Add approximately 350 ml of

1x TAE buffer to the upper buffer chamber.

Note: If you want to prerun to equilibrate the system, place the lid on the D GENE

system, attach cords, and adjust the voltage setting on the power supply.

2. Wash the wells using a 19 gauge needle and syringe with the running buffer to remove any

unpolymerized gel material in the wells before loading samples.

3. Place the D GENE lid back onto the buffer chamber. Turn the power and heater on. Adjust

the temperature setting to the desired temperature (see Section 5.1).