Hoefer SE300 miniVE User Manual
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a. Center a packing sponge on the black cathode side.
b. Lay one piece of wet filter paper on the sponge.
c. Position the equilibrated gel on the filter paper. Wet
the gel surface with a few drops of transfer buffer.
d. Lay the membrane on the gel. Do not reposition the
membrane once it contacts the gel. Use a glass rod
to roll out any air bubbles.
e. Lay one piece of wet filter paper on the membrane.
f. Lay two packing sponges on the filter paper. A
second transfer stack, if added, is placed between
these two sponges. Repeat steps b–e.
Assemble the transfer stack so that molecules will
migrate to the membrane (Fig 15).
For negatively charged macromolecules—such as
proteins run in an SDS gel and nucleic acids—
assemble the transfer stack on the black (cathode)
side. Proteins will transfer towards the red (anode) side.
Fig 15. Assembling the transfer
stack.
red = anode (+)
f
e
d
c
b
a
black = cathode (–)
Note: For best results, avoid
trapping air bubbles as each
layer is applied. Always establish
full contact along one side and
maintain contact as the layer is
lowered into position.