17
Note: do not handle the membrane without gloves.
2. Assemble the fiber pads, filter papers, gel and transfer membrane in the above
order and roll with a pipette to remove any trapped air. Place on the black and
red cassette with the membrane facing the anode (red) side, close the hinge
carefully so as to not disturb the sandwich.
3. Fill the tank with buffer solution up to the
maximum fill line
indicated on the side
of each unit. See the Appendix for recommended buffer solutions. Improved
transfer can be obtained by using chilled buffer.
Table 8. shows the volume of buffer required.
Buffer Volume
ENDURO Electroblotting
One Cassette
1380 mL
Two Cassettes
1290 mL
Three Cassettes
1200 mL
Each cooling pack takes the place of 100 mL of buffer.
Blot Running Conditions:
1. 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 in which the blot sandwiches were loaded.
2. Use of a magnetic stirring bar and plate is recommended to mix the buffer to give
consistency of transfer. A 4mm diameter stirring bar should be placed
underneath the module, in the center 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.
3. Insert the module, attach the lid and connect to a power supply.
4. Consult Table 9 for details on recommended power supply voltage settings and
blot times. Please note voltages and current will vary according to the amount of
cassettes, type and temperature of buffer and thickness and percentage of gel.
This will also affect quality of transfer so adjust the time of the blot to your
particular samples and conditions.
