Hoefer PR648 - Manual (Page 5)

Procedure for standard use of
the Hoefer PR648
The following procedure produces a well-defined
blot with sharp edges with many types of samples.
To determine the optimal load per slot for the
particular membrane and samples you are using,
we suggest first blotting a set of serial dilutions.
Setting up the Hoefer PR648
1
Place the bottom block on the lab bench. Place the
membrane support block (middle block), with the
0-ring facing down, on top of the bottom block.
The membrane recess in the membrane support block
has one curved corner. Orient the block with this
corner on the lower right (see Fig 1).
2
Wear gloves to avoid leaving fingerprints on the
membrane. Place the membrane cutting template,
ridge side down, on the membrane. Exert slight
pressure on the template to leave a mark on the
membrane. Cut the membrane along the mark. The
template has been sized to allow for swelling of all
types of membrane.
3
Soak the membrane in appropriate buffer for a few
minutes to wet it thoroughly. (A dry membrane will
draw sample under the sealing ridge and away from
the slot, resulting in a poorly defined slot image on
the membrane.)
4
Place the membrane completely within the recess,
aligning the clipped corner of the membrane with the
clipped corner of the recess.
Make sure the membrane is flat and fits completely
within the area, with edges parallel to the recess
edges. If the membrane is not positioned correctly,
lift it up and reposition it.
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