Hoefer TE70X - Manual (Page 13)

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Prepare the blotting paper
For each gel, cut at least 6 pieces of blotting paper
the same size as the gel or slightly smaller.
Gauge the thickness or number of blotting paper layers
according to the amount of buffer required; up to
300 ml of buffer may be required for larger gels or for
transfers of 60 minutes in order to prevent the stack
from drying out or the buffer from becoming depleted.
Saturate at least 3 pieces of the blotting paper with
transfer buffer. One by one, center each sheet on the
lower electrode and remove all trapped air by rolling a
clean pipet or roller from the center toward the edges.
5
Prepare the membrane
For each gel, cut 1 membrane the same size as
the gel or slightly smaller. (A larger membrane may
contact an electrode panel, creating a pathway by
which current can bypass the gel.)
Pre-wet nitrocellulose or nylon membranes with
distilled water. Pre-wet PVDF or other hydrophobic
membranes with methanol. Then soak all membrane
types in transfer buffer for 2– 5 minutes.
Note: Each 21 × 26 cm blot
paper will absorb approximately
50 ml of transfer buffer. Each 14
× 16 cm blot paper will absorb
approximately 20 ml of buffer.
Note: If not using Hoefer blotting
paper, additional pieces may be
required to achieve desired results.
Note: Always wear gloves when
handling membranes to avoid
leaving fingerprints.