Bio-Rad ReadySub-Cell GT Cells - Manual (Page 11)

5. To seal the open tray ends, engage the cam peg by turning and pressing downward simultaneously.
6. When the cam peg has dropped into the appropriate slot, turn the peg in either direction until resistance
is felt. This action seals the edges of the tray for casting.
7. Place the comb(s) into the appropriate slot(s) of the tray.
8. Prepare the desired concentration and amount of agarose in 1x electrophoresis buffer (see Section 2.1).
When the agarose solution has cooled to 50—60°C pour the molten agarose between the gates.
Warning: Hot agarose (>60°C) may cause the tray to warp or craze and will decrease the lifetime of the
tray. Warping may also result in sample wells of uneven depth.
9. Allow 20—40 min for the gel to solidify at room temperature.
10. Carefully remove the comb from the solidified gel.
11. Disengage the cam peg by turning and lifting upward. Slide the movable wall away from the tray.
Remove the tray from the Gel Caster or Mini-Gel Caster.
Note: While the gel is solidifying, a light seal is formed between the gasket and the gel (especially for low
percentage agarose gels [<0.8%]). Before moving the wall away from the tray, carefully lift the tray on
one side to release the seal or use a spatula to break the seal between the agarose and gasket.
12. Place the tray onto the leveled Sub-Cell base so that the sample wells are near the cathode (black).
DNA samples will migrate toward the anode (red) during electrophoresis.
13. Submerge the gel beneath 2 to 6 mm of 1x electrophoresis buffer (see Section 3, Gel and
Electrophoresis Reagent Preparation). Use greater depth overlay (more buffer) with increasing voltages
to avoid pH and heat effects.
Removable tray (UVTP) gel casting using tape
1. Seal the ends of the UVTP gel tray securely with strips of standard laboratory tape. Press the tape firmly
to the edges of the gel tray to form a fluid-tight seal.
2. Level the gel tray on a leveling table or workbench using the leveling bubble provided with the instrument.
3. Prepare the desired concentration and amount of agarose in 1x electrophoresis buffer (see Section 2.1).
When the agarose solution has cooled to 50—60°C pour the molten agarose into the gel tray.
Warning: Hot agarose (>60°C) may cause the tray to warp or craze and will decrease the lifetime of the
tray. Warping may also result in sample wells of uneven depth.
Engage and seal
(press down and rotate)
Movable wall
of gel caster
Fixed wall
of gel caster
Lift cam lever up
7
Fig. 2.1. Sealing the UVTP tray for gel casting.
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