Action
Step
For each gel, cut at least 6 pieces of blotting paper the same size as the gel
or slightly smaller. Make sure to cut the blotting paper to the same size as
the gel.
6
Saturate the blotting paper with transfer buffer.
7
For each gel, cut 1 membrane the same size as the gel or slightly smaller.
Note:
Always wear gloves when handling membranes to avoid leaving fingerprints.
8
Note:
A larger membrane may touch an electrode panel, creating a pathway by
which current can bypass the gel.
Pre-wet nitrocellulose or nylon membranes with distilled water. Pre-wet
polyvinylidene difluoride (PVDF) or other hydrophobic membranes with
methanol.
9
Soak all membrane types in transfer buffer for 2 to 5 minutes.
10
Assemble a transfer stack with
a single gel
Follow the instructions below to assemble the transfer stack.
Always wear gloves when handling membranes to avoid leaving fingerprints.
Note:
Action
Step
One by one, center each sheet on the lower electrode. Refer to
of a transfer stack with a single gel, on page 37
.
1
NOTICE
Stack each layer with care, with edges parallel. As each
layer is added, remove all air pockets by rolling a clean
pipet from the center to the edges. Add a few drops of
buffer to trouble areas to help remove air pockets.
Note:
For best results, the transfer stack should be centered on the electrode panels.
SEMIPHOR TRANSPHOR UNIT series Operating Instructions 29281777 AA
39
5 Operation
5.1 Assemble transfer stack
