C.B.S. Scientific EBU-222 - Manual (Page 8)

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CASSETTE ASSEMBLY
9. Remove entire assembly from pyrex dish and load into desired position (slot) in buffer chamber with black
screen facing towards black terminal (cathode) and red screen facing towards red terminal (anode).
3.3 STANDARD ELECTRO-BLOT TRANSFER
1. Fill the chamber with transfer buffer. (Buffers should be prepared fresh with reagent grade chemicals and
pre-cooled). The transfer buffer should come up to the top of the platinum labyrinth. The buffer should not
come in contact with the banana plugs when the gel cassette sandwich(es) are immersed in the unit.
2. Align safety cover over the unit and carefully attach. Begin cycling of coolant.
3. Connect the leads to the power supply, matching the color-coded red to red and black to black. See Section
4.1 for recommended power conditions.
4. Transfer times will vary according to several parameters. Optimization of electro-blotting transfers must be
determined empirically. Keep in mind the following principles that govern the movement of molecules of gel
electrophoresis:
• Thicker or higher percentage gels will take longer to transfer than thinner or lower percentage gels.
• Large molecules will need extended transfer times to completely transfer.
• Actual transfer times for defined conditions can be approximated by running molecular weight standards.
3.4 REMOVING THE CASSETTES
1. Turn the power supply off and disconnect the leads from the power supply.
2. Remove the safety cover from the unit, by placing thumbs on white posts next to red & black connectors,
then pushing down while pulling up with fingers under lid. DO NOT pull on power cords.
3. Gently lift the cassette from the unit. Always wear gloves, eye protection and protective clothing if buffer and/
or gel contain Ethidium Bromide. Ethidium Bromide is a powerful mutagen; gloves, eye protection and pro-
tective clothing should always be worn when handling the gel or buffer solutions. See Material Data Safety
Sheets.
4. Mark the orientation of the membrane with a pencil or by cutting off a corner and take apart the cassette
carefully.
5. Process membrane according to type of transfer and manufacturer’s recommendations.