Bio-Rad Foresight™ Chromatography Columns, Prepacked - Manual (Page 28)

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Recommended settings for vacuum filtration
are 1 min at –1” Hg. If liquid is still visible in
the wells of the media plate, repeat the cycle.
Recommended settings for centrifugation are
1–2 min at 300 g (or more if visual inspection
indicates residual liquid).
• The collection plate should be large
enough to hold the storage solution.
Remember to empty the collection plate
when necessary
3.6.2 Plate Equilibration
Equilibrate the desired wells using 300 μl of
buffer per well. Perform this procedure three
times. During each equilibration, incubate the
media plate on a microplate shaker for
1–2 min at 1100 rpm. Remove the
equilibration buffer at the end of each
equilibration step using either vacuum filtration
or centrifugation.
• Firmly fix the media and collection plate to
each other and to the microplate shaker
• Usually three equilibration cycles are
enough to equilibrate the media. To
monitor the progress, a micro pH probe
can be used
Note: Remember to keep the media plate on
top of a collection plate at all times.
After the last equilibration cycle, blot the
media plate on a paper towel. Blotting
eliminates droplets that might still be hanging
on the drip directors, which will lead to
non-reproducible dilution of the feed solution
flowthrough. Blotting is recommended after
the end of the sample incubation, washing,
elution, and regeneration cycles.