Consort EV1300, Manual

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EVS1000 Blot units
3
Electroblotting
Setting up the blot sandwich
Each blot sandwich should be set up as follows:
•
Cassette clamp [-ve] (black) side placed in a tray or other suitable surface.
•
Pre-soaked fibre pad (note two can be used with thin gels).
•
Two pieces of thick filter paper, about 2...3 mm thick, pre-soaked in buffer.
•
Gel.
•
Transfer membrane. Usually this requires pre-soaking but consult the manufacturer’s
instructions for the type of membrane you are using. This should be smoothed so that no air
bubbles have been trapped.
•
Two pieces of thick filter paper, about 2...3 mm thick, pre-soaked in buffer.
•
Pre-soaked fibre pad (note two can be used with thin gels).
•
Cassette clamp [+ve] (red) side slotted into the groove in the bottom of the black cassette.
Close the hinge carefully so as to not disturb the sandwich.
Fill the tank with buffer solution up to the maximum fill line indicated on the side of each unit.
Improved transfer can usually be obtained by using chilled buffer.
Buffer Volume EVS1100 EVS1200 EV1300
One cassette 1380 ml 2800 ml 5600 ml
Two cassettes 1290 ml 2620 ml 5240 ml
Three cassettes 1200 ml 2440 ml 4880 ml
Four cassettes 1110 ml 2260 ml 4520 ml
Each cooling pack will take the place of 100 ml of buffer for EVS1100 and 500 ml of buffer for
EVS1200 and EVS1300.
Blot run conditions
Insert the cassettes into the slots in the module with the black side of each adjacent to the negative
electrode. It is a good idea to note the orientation and order the blot sandwiches were loaded in. This
can be done by noting which samples were loaded adjacent to each electrode.
Use of a magnetic stirring bar and plate is recommended to mix the buffer to give consistency of
transfer. A 4 mm diameter stirring bar should be placed underneath the module, in the centre of the
tank. The cooling pack provided, pre-frozen, can be inserted at the side or front of the tank for
extended blots. Additional cooling packs can be purchased as accessories to further aid cooling.
Insert the module, fit the lid and connect to a power supply.
Consult below table for details on recommended power supply voltage settings and blot times. Please
note voltage and current will vary according to the amount of cassettes, type and temperature of