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See Also: Equipment List
- Prepare all necessary solutions.
- Cut nitrocellulose transfer membrane and 6 sheets of GB004 gel blot paper to the EXACT size of the polyacrylamide gel.
- Equilibrate gel in cathode transfer buffer for EXACTLY 5 minutes.
- Prewet the nitrocellulose transfer membrane in anode transfer buffer for 5 minutes.
- Wipe the surface of the anode (in the base of the blotter) with a paper towel that has been saturated with anode buffer.
- Prewet 3 sheets of GB004 gel blot paper until saturatedwith anode buffer and place them on the anode.
- Place nitrocellulose transfer membrane (prewet) on top of the saturated stack of gel blot paper.
- Remove all air bubbles by gently rolling a glass pipet or stirring rod over the stack (EXTREMELY IMPORTANT!!!)
- Place the equilibrated prewet gel in the transfer membrane, taking care not to create air bubbles. (EXTREMELY IMPORTANT!!!)
- Saturate 3 sheets of GB004 gel blot paper in cathode buffer and place them on top of the gel, removing air bubbles completely.
- Wipe the surface of the cathode (in the top) with a paper towel saturated with cathode buffer.
- Place the lid (cathode) on the base (anode) lining up the red and black rods with the holes in the lid.
- Connect the black cable on the Pronto lid to the hole containing the (-) electrode.
- Connect one end of the red cable to the hole containing the (+) electrode on the lid
- Connect both the red and black cables to the power supply [black = (-) and red = (+)].
- Plug in and turn on the power supply
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