Bio-Rad Criterion™ TBE-Urea Precast Gels - Manual (Page 40)

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Foam pad
Membrane
Gel
Filter paper
Filter paper
Foam pad
4. Place the assembled cassette into the groove in the tank, aligning the red side of the card with the
red electrode. Make sure the magnetic stirbar is free to move. Repeat steps 2–4 for a second blot.
5. Add the remaining transfer buffer to the fill level marked on the tank, place the tank on a stir plate,
and begin stirring to maintain even buffer temperature and ion concentration during the transfer.
6. Connect the Criterion blotter to a PowerPac ™ HC power supply and begin transfer.
For many proteins, excellent transfer efficiency is obtained in 30 min at a constant voltage of 100 V. For
best results, optimize conditions for proteins of interest. Large proteins (>150 kD) may take 60 min, while
smaller proteins (<30 kD) may transfer in 20 min. Refer to the Criterion Blotter Instruction Manual (bulletin
4006190) or the Protein Blotting Guide (bulletin 2895) for additional information.
13.2.3 Transfer Using the Trans-Blot ® Turbo ™ System
1. Open the transfer pack and assemble the components on the cassette in the order shown. For best
results, use the roller to remove any air trapped between the layers. If using a single mini or midi
sandwich, place the sandwich in the middle of the cassette bottom. With two mini gels, place the
sandwiches on a midi stack with the foot of each gel facing the center of the stack.
2. Place the lid on the cassette and lock the lid in place by turning the knob clockwise, using the
symbols on the lid as a guide. Slide the cassette into the appropriate bay of the Trans-Blot Turbo cell.
Each cassette and bay can hold up to two mini gels or one midi gel (Table 13.1).
3. Start the transfer. With the cassette in the cell, press TURBO and select the gel type. Press A:RUN
or B:RUN to begin the transfer. Press LIST to select a Bio-Rad optimized protocol (Table 13.2) or a
user-defined protocol. Press NEW to create and run a new protocol.
4. When transfer completes, RUN COMPLETE appears. Pull the cassette straight out of the slot and
unlock the lid. Disassemble the blotting sandwich.
b) Open the cassette and place it in the back (large) compartment of the tray so the red plate
with the handle is vertical (anode) and the black plate (cathode) is horizontal and submerged in
transfer buffer.
c) Assemble the sandwich as shown, placing the gel closest to the black side of the cassette and
the membrane closest to the red plate. Use a blot roller to remove air trapped between the
layers of the blot assembly.
Assembly of the gel blot sandwich with the Criterion blotter .
Criterion Precast Gels