C.B.S. Scientific EBU-102P - Manual (Page 11)

C.B.S. œ Scientific
11
Electro-Blotting System
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 protective 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.