C.B.S. Scientific EBU-204 - Manual (Page 10)

10
EBU-204 Mini-Blotting System Instructions 6-2-14
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