Hoefer SE300 miniVE - Manual (Page 28)

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Prepare the transfer stack
Transfer the sample as soon as possible after
electrophoresis to minimize sample diffusion
within the gel. Electrophoretic transfer can
be performed on as many as four mini gels at
one time, if two gels are placed in each of two
modules.
The transfer stack consists of the gel and
membrane, filter paper, and three packing
sponges. The gel determines the size of the
membrane and filter paper.

For each gel, cut the membrane and two pieces of
filter paper the same size as the gel, but no larger
than 8.5 × 10.5 cm.

Equilibrate the gel in transfer buffer for 10 minutes.
Equilibration allows the gel to swell or shrink before it
contacts the transfer membrane and removes excess
buffer salts and detergents from the gel. Longer equil-
ibration may result in diffuse bands.

Pre-wet nitrocellulose or nylon membranes in distilled
water, taking care not to trap air bubbles.
Dip one end of the membrane into the buffer and
slowly submerge it, allowing it to wet by capillary
action.
Pre-wet PVDF or other hydrophobic membranes in
methanol.
After pre-wetting, soak all membrane types in transfer
buffer for 2– 5 minutes.

Wet the two pieces of filter paper in transfer buffer.
Important! Try to place the gel
correctly the first time. Proteins
may begin to transfer immediately.
Once transfer begins, moving the
gel will distort results or cause
“shadow bands” on the blot.