Bio-Rad ReadySub-Cell GT Cells - Manual (Page 10)

5. Boil and swirl the solution until all of the small translucent agarose particles are dissolved. With the small
flask still in place, set aside to cool to 60°C before pouring.
2.2 Casting Agarose Gel Slabs
There are several ways to cast agarose submarine gels using the Sub-Cell GT systems. Gels may be
cast with a UV-transparent plastic (UVTP) tray directly on the gel stage of the Sub-Cell GT bases using the
gel casting gates. Gels may also be cast on the removable UVTP trays with the aid of the gel caster or with
standard laboratory tape.
Casting gels on the base stage with the UVTP tray
1. Level the cell using the leveling bubble provided.
2. Place the UVTP tray on the gel stage.
Note: The Mini-Sub Cell GT requires the 7 x 7 cm UVTP tray for casting in the GT base. The Wide-Mini-
Sub Cell GT requires the 15 x 7 cm UVTP tray and the Sub-Cell GT system requires the 15 x 15 cm
UVTP tray for casting in the GT base.
3. Slide the gel casting gates into the slots at opposite ends of the GT gel stage. Insure that the gates are
evenly seated in the slots and the gates uniformly contact all edges of the UVTP tray. The weight of the
gates provides a tight seal to prevent any leakage problems during gel casting*.
* Note: If leakage occurs while pouring the gel on the casting tray atop the stage, chill the casting gates
in the freezer for 2-3 minutes. Place the casting gates into the slots when ready to pour the gel. The
chilled casting gates will prevent the gel solution from leaking out of the tray and into the chambers.
4. Place the comb(s) into the appropriate slot(s) of the trays so that the sample wells are near the cathode
(black). DNA samples will migrate toward the anode (red) during electrophoresis.
5. 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.
6. Allow 20—40 minutes for the gel to solidify at room temperature.
7. Carefully remove the comb from the solidified gel. Remove the gel casting gates.
8. 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 prevent pH and heat effects.
Removable tray (UVTP) gel casting using a Gel Caster or Mini-Gel Caster
1. Level the Gel Caster or Mini-Gel Caster using the leveling feet in the gel caster and the leveling bubble
provided.
2. Disengage and slide the movable wall to the open end of the Gel Caster or Mini-Gel Caster by turning
and lifting the cam peg upward.
Note: If casting more than one gel with the Gel Caster, add the removable gel casting wall to the gel
caster. The removable wall will allow casting using two 15 x 10 cm trays, four 7 x10 cm trays or one
15 x 10 cm and one 15 x15 cm trays.
3. Place the open edge of the UVTP tray against the fixed wall of the Gel Caster or Mini-Gel Caster.
4. Slide the movable wall against the edge of the UVTP tray (Figure 2.1).
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