4
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.
um TE70X-IMA0
•
p6
Note: Each 21 × 26 cm blot paper
will absorb approximately 50 ml
of transfer buffer. Each 14
× 16
cm blot paper will absorb approxi
-
mately 20 ml of buffer.
Note: Always wear gloves when
handling membranes to avoid
leaving fingerprints.
To Next Page
Loading...