Bio-Rad Bio-Gel P Polyacrylamide Gel - Manual (Page 8)

6. Pour the even slurry into the column in a single, smooth move-
ment. Avoid splashing the slurry, to insure even packing, and to
avoid trapping air bubbles (Figure 6).
7. When a 2-5 cm bed has formed, allow column to flow until the
column is packed.
8. When the column is packed, close the column outlet and insert the
flow adaptor. Open the column outlet and pass 2 bed volumes
of buffer through the column at the operating flow rate.
9. Close the outlet and adjust the flow adaptor down to the level of
the gel bed. Load sample onto the upper bed surface by pumping
or injecting sample onto the gel bed through the flow adaptor. If
sample is injected, the injection flow rate should not exceed the
recommended elution flow rate.
10. If a flow adaptor is not to be used, remove excess gel to the
desired bed height once the column is packed and attach column
to a reservoir. Pass 2 bed volumes of buffer through the column
at the operating flow rate. Drain the buffer down to the level of
the gel bed and layer the sample carefully onto the upper bed
surface, allowing it to drain into the bed. Follow this with addi-
11 10
5. Affix a funnel to top of column, close column exit, and add
enough buffer to fill 20% of column (Figure 5).
Fig. 5. Fig. 6.
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