Bio-Rad Bio-Gel P Polyacrylamide Gel - Manual (Page 7)

2. Allow Bio-Gel P-2 through P-10 gels to hydrate 4 hours at room
temperature (1 hour if buffer was previously brought to 100 °C
and then allowed to cool after addition of gel). Bio-Gel P-30
through P-100 gels will require 12 hours at 20 °C, or 4 hours
starting at 100 °C. After initial uniform suspension of beads is
established, it is not necessary to stir; let settle during hydration
(Figure 2).
3. After hydration is complete, decant half of supernatant (Figure 3).
Transfer the solution to a filter flask and attach to a vacuum
source. Degas the solution for 5-10 minutes with occasional
swirling of the flask (Figure 4). Do not use a stir bar, as it may
damage the gel.
4. Add two bed volumes of degassed buffer and swirl gel gently.
Allow gel to settle until 90-95% of the particles have settled.
Decant or remove supernatant by suction to remove fines. Repeat
up to 4 times to remove > 90% of the fines.
9 8
3.3 Preparation of the Gel
1. Gradually add dry Bio-Gel P media to buffer in a beaker. The
amount of Bio-Gel P gel required to pack a column of known
volume may be estimated by using the hydrated bed volume
given in Table 2. Allow for gel loss during handling. Use twice
as much buffer as the expected packed bed volume (Figure 1).
Fig. 1. Fig. 2.
Fig. 3. Fig. 4.
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