Bio-Rad Trans-Blot® Turbo™ Transfer System - Manual (Page 22)

Once the stacks are positioned in the cassette base, place the cassette lid on the base. The lid is
reversible, but ensure that the electrical contacts fit closely into the slots in the base. Press the lid down
firmly and turn the dial clockwise to engage the lid pins into the locking slots.
Slide the cassette (with the dial facing up) into one of the Trans-Blot Turbo instrument bays until it makes
contact with the magnetic interlock in the back of the instrument tub and you hear a click. The cassette
can be inserted into the bays with or without power to the system.
The bays are labeled A (top) and B (bottom), and cassettes can be inserted interchangeably into the
bays. There is no requirement for both cassettes to be inserted; one bay (either A or B) can be left empty
when a protocol is run.
The instrument is now ready to begin a transfer protocol. Refer to the following sections for details on
transfer using preprogrammed and user-defined protocols.
Note: A transfer for one cassette can be started while you are assembling a second cassette. The
second transfer can be started independently as long as the same protocol is being used for both.
Fig. 11. Suggested placement of assembled transfer stacks in a cassette. a. Midi stack and gel
placement. b. Mini stack and gel placement. c. Two mini gels on a midi stack.
Set the transfer stack close to the center of the cassette. A midi stack only fits along the long axis of
the cassette. It is not necessary to move an assembled transfer pack if it is only slightly off center in the
cassette.
a. b. c.
3.3 Transfer Using RTA Transfer Kits
Trans-Blot Turbo RTA transfer kits provide the same reagents and consumables available in the transfer
packs, but in a ready-to-assemble format. The protocol for using them is similar to that used for the
transfer packs, except membranes and transfer stacks must be soaked in Trans-Blot Turbo transfer
buffer before use. Do not equilibrate the gel before transfer.
1. Prepare the Trans-Blot Turbo Transfer Buffer according to instructions on the bottle.
2. Wet and equilibrate the membrane and transfer stacks:
• Nitrocellulose membrane — immerse in 30 ml of 1x transfer buffer and equilibrate for 2–3 min
• PVDF and LF PVDF membranes — immerse in 100% methanol or ethanol (use reagent grade
~85% or molecular biology grade ~95%–98% purity) until the membrane is translucent. Transfer
to a soaking tray containing 30 ml of 1x transfer buffer, ensure the membrane is submerged, and
equilibrate 2–3 min
• Transfer stacks —
• Midi stacks — immerse two stacks seperated by blue sheets in two soaking each containing
50 to 70 ml of transfer buffer for 2–3 min
• Mini stacks — immerse two stacks seperated by blue sheets side-by-side in a soaking tray
containing 50 to 70 ml of transfer buffer for 2–3 min
2. Follow the procedure described in Figures 8-10 above for assembling the transfer sandwiches in
transfer cassettes, followed by an addtional step: once assembled, remove excess transfer buffer by
inverting the cassette base with the assembled stack carefully held in place into a waste container.
Once the excess buffer is removed, place the cassette lid on the base, and proceed with the
transfer step (3.4).
10 Tech Support: 1-800-424-6723 • www.bio-rad.com
Trans-Blot® Turbo™ Blotting System