Bio-Rad Trans-Blot® SD Semi-Dry Transfer Cell - Manual (Page 6)

Section 4
General Assembly of Unit for Transfer
1. Saturate two pieces of precut Extra Thick Blot Paper (provided with the kit) and precut
Zeta-Probe ® GT membrane (also provided with the kit) or any other transfer membrane
in 0.5x TBE buffer. Equilibrate the transfer membrane for at least 10 minutes. The trans-
fer membrane and blot paper must have the same dimensions as the gel for proper trans-
fer to occur. If necessary, cut the transfer membrane and the Extra Thick Blot Paper to the
required dimensions with a clean razor blade or scissors. Precut Zeta-Probe GT blotting
membrane and extra thick blot paper can be purchased separately.
2. To assemble the transfer sandwich, hold up one piece of blot paper, allow the excess
buffer to drain off, and lay it flat on the platinum anode. Using a clean pipette, roll out any
air bubbles that may be trapped under the blot paper with a top-to-bottom and left-to-
right rolling motion.
3. Place the equilibrated transfer membrane on top of the blot paper and roll out the air bub-
bles.
4. Carefully place the agarose gel on top of the membrane, well side up. Make sure all the
edges are aligned and air bubbles are rolled out.
5. Take the other piece of wetted blot paper, again allowing the excess buffer to drain off,
and place it on top of the agarose gel. Roll out the blot paper to remove air bubbles and
add approximately 15 ml of transfer buffer (0.5x TBE) on top to resaturate the sandwich.
Be sure to remove any excess buffer that is present on the anode electrode surface.
6. Place the support frame around the gel/membrane/blot paper stack and connect the cath-
ode electrode by locking it into place without disturbing the stack. The frame accomodates
a 6 mm thick gel, transfer membrane, and two pieces of the Extra Thick Blot Paper pro-
vided with the kit and available from Bio-Rad.
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